Crystalline form of 6-[(4s)-2-methyl-4-(2-naphthyl)-1,2,3,4-tetrahydroisoquinolin-7-yl]pyridazin-3-amine

ABSTRACT

The present disclosure generally relates to a crystalline form of 6-[(4S)-2-methyl-4-(naphthyl)-1,2,3,4-tetrahydroisoquinolin-7-yl]pyridazin-3-amine. The present disclosure also generally relates to pharmaceutical compositions comprising the crystalline form, as well of methods of using a crystalline form in the treatment of depression and other conditions and methods for obtaining such crystalline form.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of application Ser. No. 12/995,771filed Dec. 2, 2010 which is the 371 National Stage of InternationalApplication No. PCT/US2009/046256 filed Jun. 4, 2009 which claims thebenefit under 35 U.S.C. §119(e) of 61/058,717 filed Jun. 4, 2008.

The present disclosure generally relates to a crystalline form of6-[(4S)-2-methyl-4-(naphthyl)-1,2,3,4-tetrahydroisoquinolin-7-yl]pyridazin-3-amine.The present disclosure also generally relates to a pharmaceuticalcomposition comprising a crystalline form, as well of methods of using acrystalline form in the treatment of depression and other conditions andmethods for obtaining such crystalline form.

Major depression (unipolar depression) is a condition of high prevalenceand very high global burden. The prevalence of the disease in the US wasestimated at over 6% in a twelve month period and 16% over a lifetime(See, Kessler R. C. et al., “Prevalence, severity, and comorbidity of12-month DSM-IV disorders in the National Comorbidity SurveyReplication”, Arch. Gen. Psychiatry, 62:617-627 (2005)). According tothe World Health Organization, the impact of the disorder is high withdepression being the fourth leading cause of disease burden (See, Ustun,J. L. et al., “Global burden of depressive disorders in the year 2000”,Brit. J. Psychiatry, 184:386-392 (2004)). There are many symptomsassociated with major depression. These symptoms generally fall into thetwo major categories of depressed mood and loss of interest and pleasure(anhedonia).

While the etiological basis of mood disorders is not clear, there arelines of evidence that indicate that the disorders are associated with aheterogeneous dysregulation of monoaminergic systems, i.e., thoseinvolving dopamine, norepinephrine and serotonin (See, Garlow, S. J. etal., “The neurochemistry of depressive disorders: clinical studies”,Neurobiology of Mental Illness, 2nd Ed., Charney, D. S. et al., eds.,pp. 440-460 (2004)). For instance, abnormalities in levels of serotonin,norepinephrine and dopamine metabolites are commonly reported indepressed patients. Furthermore, the leading pharmacotherapies appliedto depression are aimed at monoamine systems, primarily serotonin andnorepinephrine.

The response rate of typical antidepressants is reported to be on theorder of 65% while remission from the disorder is only about 30% (See,Hirschfeld, R. M. et al., “Partial response and nonresponse toantidepressant therapy: current approaches and treatment options”, J.Clin. Psychiatry, 63:826-837 (2002)). The onset of action of drugs fordepression is often long, on the order of 4 weeks to response.Accordingly, new drugs for treating depression, as well as otherconditions, that improved response rate, remission or onset wouldrepresent an important improvement over current therapies. The spectrumof symptom relief is also an important component to efficacy sinceselective serotonin reuptake inhibitors (SSRIs) andserotonin-norepinephrine reuptake inhibitors (SNRIs), while effective onsymptoms related to depressed mood, are typically not optimallyeffective in the areas of motivation and interest in pleasure.

Accordingly, there is a need for increased efficacy over SSRIs andSNRIs. In addition, reduction in the degree of side effects is alsodesired. For example, addressing decreased libido as well as sleep andgastrointestinal disorders (GI) disorders associated with currentanti-depressants would provide substantial benefits to patients.

One strategy that has emerged in antidepressant development is the useof triple reuptake inhibitors (TRUIs), which jointly inhibit reuptake ofserotonin, norepinephrine, and dopamine These compounds have beenhypothesized to have a more rapid onset of activity and better efficacyover single or dual reuptake inhibitor antidepressants in part due tothe addition of the dopamine component. While the monoamine theory ofdepression is centered around norepineprine and serotonin, there aredata that suggest the involvement of dopamine as well (See, Dunlop, B.W. et al., “The role of dopamine in the pathophysiology of depression”,Arch. Gen. Psychiatry, 64:327-337 (2007) and Papakostas, G. I. et al.,“A metaanalysis of clinical trials comparing moclobemide with selectiveserotonin reuptake inhibitors for the treatment of major depressivedisorder”, Can. J. Psychiatry, 51:783-790 (2006)) Motivation,concentration and the ability to experience pleasure are regulated inpart through central dopaminergic systems and they are also negativelyimpacted in depression. Subgroups of depressed patients have also beenobserved to have decreased cerebrospinal fluid (csf) levels of thedopamine metabolite homovanillic acid, suggesting decreased dopaminergicfunction in those patients. Over the past several years, a number ofstudies have demonstrated the benefit of combining drugs that havedopamine transporter (DAT) inhibition properties, like bupropion andmethylphenidate, with SSRIs or SNRIs (See, Marshall, R. D. et al.,“Paroxetine/bupropion combination treatment for refractory depression”,16:80-81 (1996); Bodkin, J. A. et al., “Combining serotonin reuptakeinhibitors and buproprion in partial responders to antidepressantmonotherapy”, J. Clin. Psychiatry, 58:137-145 (1997); Lavretsky, H. etal., “Combined treatment with methylphenidate and citalopram foraccelerated response in the elderly: an open trial”, J. Clin.Psychiatry, 64:1410-1414 (2003); Trivedi, M. H. et al., “Medicationaugmentation after the failure of SSRIs for depression”, N. Engl. J.Med., 354:1243-1252 (2006)).

It is desired to provide drugs that target the DAT, serotonintransporter (SERT), and norepinephrine transporter (NET). Desirably, thedrug would provide an optimal ratio of SERT, DAT and NET inhibition.Accordingly, SERT, DAT and NET occupancies are important pharmacologicalcriteria for consideration. In one aspect of the invention, compoundsare provided that lead to about 20-60% DAT occupancy while maintainingSERT occupancy greater than about 80%.

In accordance with the present disclosure, a particular crystalline formof6-[(4S)-2-methyl-4-(naphthyl)-1,2,3,4-tetrahydroisoquinolin-7-yl]pyridazin-3-amineis provided that may be useful in treating depression in addition to avariety of other conditions, e.g., anxiety disorders, pain, attentiondeficit disorder (ADD), smoking cessation and obesity.

For purposes of clarification, the free base racemate ofrac-6-[(4)-2-methyl-4-(naphthyl)-1,2,3,4-tetrahydroisoquinolin-7-yl]pyridazin-3-amine,represented by Formula (I) is referred to herein as Compound (I). The(4S)-enantiomer of Compound 1, represented by Formula (II), is referredto herein, in general (without reference to any specific crystallineform), as Compound 2. The crystalline form of Compound 2 in accordancewith the present disclosure, also represented by Formula (II), isreferred to herein as Form N-1.

It has been found that Form N-1, can be repeatedly crystallized andprovide high aqueous solubility and excellent purification capacity,thereby making it a suitable candidate for drug development.

In its first aspect, the present disclosure provides Form N-1.

In a second aspect, the present disclosure provides Form N-1,characterized by the following unit cell parameters:

Cell dimensions:

a=8.4299(4) Å

b=6.0698(3) Å

c=19.0689(12) Å

alpha=90°

beta=100.169(2)°

gamma=90°

Space group Monoclinic, P2₁

Volume 960.39(9) Å³

Z, Calculated density 2, 1.267 g/cm³

wherein measurement of free base crystalline form is at a temperaturebetween about 20° C. to about 25° C.

In a third aspect, the present disclosure provides Form N-1,characterized by fractional atomic coordinates within the unit cell aslisted in Table 3, Atomic Coordinates.

In a fourth aspect, the present disclosure provides Form N-1 withcharacteristic peaks in the powder X-ray diffraction pattern at valuesof two theta of 4.6±0.1, 9.4±0.1, 10.6±0.1, 14.1±0.1, 15.4±0.1,18.2±0.1, 19.5±0.1 at a temperature between about 20° C. and about 25°C., based on a high quality pattern collected with a diffractometer(CuKα) with a spinning capillary with 2θ calibrated with a NationalInstitute of Standards and Technology (NIST) or other suitable standard.

In a fifth aspect, the present disclosure provides Form N-1characterized by a melt with decomposition endotherm with onsettypically in the range of 235-245° C.

In a sixth aspect, the present disclosure provides substantially pureForm N-1.

In a seventh aspect, the present disclosure provides pharmaceuticalcompositions comprising Form N-1 and a pharmaceutically acceptablecarrier or diluent.

In an eighth aspect, the present disclosure provides pharmaceuticalcompositions comprising Form N-1 in combination with one or moreadditional compounds having anti-depression activity.

Other aspects of the present disclosure may comprise suitablecombinations of two or more of embodiments and/or aspects disclosedherein.

Further, other aspects of the disclosure will be apparent according tothe description provided below.

FIG. 1 illustrates experimental and simulated powdered X-ray diffractionpatterns (CuKα λ=1.54178 Å at T=room temperature) of Form N-1.

FIG. 2 illustrates the differential scanning calorimetry pattern of FormN-1.

FIG. 3 illustrates the Solid State Nuclear Magnetic Resonance (SSNMR)spectrum of Form N-1.

DEFINITIONS

Stereochemical definitions and conventions used herein generally followMcGraw-Hill Dictionary of Chemical Terms, Parker, S. P., ed.,McGraw-Hill Book Company, New York (1984) and Eliel, E. et al.,Stereochemistry of Organic Compounds, John Wiley & Sons, Inc., New York(1994). Many organic compounds exist in optically active forms, i.e.,they have the ability to rotate the plane of plane-polarized light. Indescribing an optically active compound, the prefixes D and L or R and Sare used to denote the absolute configuration of the molecule about itschiral center(s). The prefixes d and l or (+) and (−) are employed todesignate the sign of rotation of plane-polarized light by the compound,with (−) or 1 meaning that the compound is levorotatory and (+) or d,meaning the compound, is dextrorotatory. For a given chemical structure,these compounds, called stereoisomers, are identical except that theyare mirror images of one another. A specific stereoisomer of a mirrorimage pair may also be referred to as an enantiomer, and a mixture ofsuch isomers is often called an enantiomeric mixture.

The term “chiral” refers to molecules which have the property ofnon-superimposability of the mirror image partner, while the term“achiral” refers to molecules which are superimposable on their mirrorimage partner.

The term “derivative” means a chemically modified compound wherein themodification is considered routine by the ordinary skilled chemist, suchas an ester or an amide of an acid, protecting groups, such as a benzylgroup for an alcohol or thiol, and tert-butoxycarbonyl group for anamine.

The term “enantiomers” refers to two stereoisomers of a compound whichare non-superimposable mirror images of one another.

The term “halogen” as used herein and in the claims is intended toinclude fluorine, bromine, chlorine and iodine while the term “halide”is intended to include fluoride, bromide, chloride and iodide anion.

The term “patient” includes both human and other mammals.

The term “pharmaceutical composition” means a composition comprisingForm N-1 in combination with at least one additional pharmaceuticalcarrier, i.e., adjuvant, excipient or vehicle, such as diluents,preserving agents, fillers, flow regulating agents, disintegratingagents, wetting agents, emulsifying agents, suspending agents,sweetening agents, flavoring agents, perfuming agents, antibacterialagents, antifungal agents, lubricating agents and dispensing agents,depending on the nature of the mode of administration and dosage forms.Ingredients listed in Remington's Pharmaceutical Sciences, 18th Ed.,Mack Publishing Company, Easton, Pa. (1999) for example, may be used.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complicationcommensurate with a reasonable risk/benefit ratio.

The term “pharmaceutically acceptable salt” is intended to includenontoxic salts synthesized from a compound which contains a basic oracidic moiety by conventional chemical methods. Generally, such saltscan be prepared by reacting the free acid or base forms of thesecompounds with a stoichiometric amount of the appropriate base or acid,respectively, in water or in an organic solvent, or in a mixture of thetwo; generally, nonaqueous media like ether, ethyl acetate, ethanol,isopropanol, or acetonitrile are preferred. Lists of suitable salts arefound in Remington's Pharmaceutical Sciences, 18th Ed., Mack PublishingCompany, Easton, Pa., p. 1445 (1990). Suitable inorganic bases such asalkali and alkaline earth metal bases include The term “polymorph”refers to crystalline forms having the same chemical composition butdifferent spatial arrangements of the molecules, atoms, and/or ionsforming the crystal.

The terms “racemic mixture” and “racemate” refer to an equimolar mixtureof two enantiomeric species, devoid of optical activity. In addition, asused herein, the terms “racemic mixture” and “racemate” are intended toinclude equimolar mixtures of two enantiomers.

The term “solvate” means a physical association of a compound of thisdisclosure with one or more solvent molecules. This physical associationincludes hydrogen bonding. In certain instances the solvate will becapable of isolation, for example when one or more solvent molecules areincorporated in the crystal lattice of the crystalline solid. “Solvate”encompasses both solution-phase and isolable solvates. Exemplarysolvates include hydrates, ethanolates, methanolates, and the like.

The term “stereoisomers” refers to compounds which have identicalchemical constitution, but differ with regard to the arrangement of theatoms or groups in space.

The term “substantially pure” refers to chemical purity and form purity.More specifically, substantially pure Form N-1 comprises at least about95 wt %, preferably at least about 98 wt %, more preferably at leastabout 99 wt % of Form N-1 and less than about 5 wt %, preferably lessthan about 2 wt %, and more preferably less than about 1 wt % of othercompounds having a different chemical structure than Compound 2.Additionally, substantially pure Form N-1 comprises at least about 95 wt%, preferably at least about 98 wt %, more preferably at least about 99wt % of Form N-1 and less than about 5 wt %, preferably less than about2 wt %, and more preferably less than about 1 wt % of any othercrystalline form of Compound 2. This means that the Form N-1 preferablycontains less than about 5 wt % of other compounds, and less than about5 wt % of any other form (also referred to as “phase homogenicity”).

The term “therapeutically effective amount” means the total amount ofForm N-1 that is sufficient to show a meaningful patient benefit. Whenapplied to an individual active ingredient, administered alone, the termrefers to that ingredient alone. When applied to a combination, the termrefers to combined amounts of the active ingredients that result in thetherapeutic effect, whether administered in combination, serially orsimultaneously. If Form N-1 is used in combination with anothermedication, i.e., drug, the combination of compounds described hereinmay result in a synergistic combination. Synergy, as described forexample by Chou et al., Adv. Enzyme Regul., 22:27-55 (1984), occurs whenthe effect of the compounds when administered in combination is greaterthan the effect of the compounds when administered alone as singleagents.

The term “treating” refers to: (i) preventing a disease, disorder orcondition from occurring in a patient which may be predisposed to thedisease, disorder and/or condition but has not yet been diagnosed ashaving it; (ii) inhibiting the disease, disorder or condition, i.e.,arresting its development; and (iii) relieving the disease, disorder orcondition, i.e., causing regression of the disease, disorder and/orcondition.

General Preparation of Crystalline Materials

Crystalline forms may be prepared by a variety of methods, including forexample, crystallization or recrystallization from a suitable solvent,sublimation, growth from a melt, solid state transformation from anotherphase, crystallization from a supercritical fluid, and jet spraying.Techniques for crystallization or recrystallization of crystalline formsfrom a solvent mixture include, for example, evaporation of the solvent,decreasing the temperature of the solvent mixture, crystal seeding asupersaturated solvent mixture of the molecule and/or salt, freezedrying the solvent mixture, and addition of antisolvents(countersolvents) to the solvent mixture. High throughputcrystallization techniques may be employed to prepare crystalline formsincluding polymorphs. Crystals of drugs, including polymorphs, methodsof preparation, and characterization of drug crystals are discussed inByrn, S. R. et al., Solid-State Chemistry of Drugs, 2nd Ed., SSCI, WestLafayette, Ind. (1999).

For crystallization techniques that employ solvent, the choice ofsolvent or solvents is typically dependent upon one or more factors,such as solubility of the compound, crystallization technique, and vaporpressure of the solvent, or the ability to afford a substantially purecrystalline form. Combinations of solvents may be employed, for example,the compound may be solubilized into a first solvent to afford asolution, followed by the addition of an antisolvent to decrease thesolubility of the compound in the solution and to afford the formationof crystals. An antisolvent is a solvent in which the compound has lowsolubility.

In one method to prepare crystals, a compound is suspended and/orstirred in a suitable solvent to afford a slurry, which may be heated topromote complete or partial dissolution. The term “slurry”, as usedherein, means a saturated solution of the compound, which may alsocontain an additional amount of the compound to afford a heterogeneousmixture of the compound and a solvent at a given temperature.

Seed crystals may be added to any crystallization mixture to promotecrystallization. Seeding may be employed to control growth of aparticular polymorph or to control the particle size distribution of thecrystalline product and/or afford a substantially pure crystalline form.Accordingly, calculation of the amount of seeds needed depends on thesize of the seed available and the desired size of an average productparticle as described, for example, in Mullin, J. W. et al., “ProgrammedCooling of Batch Crystallizers,” Chemical Engineering Science,26:369-377 (1971). In general, seeds of small size are needed to controleffectively the growth of crystals in the batch. Seed of small size maybe generated by sieving, milling, or micronizing of large crystals, orby micro-crystallization of solutions. Care should be taken that millingor micronizing of crystals does not result in any change incrystallinity of the desired crystal form (i.e., change to amorphous orto another polymorph).

A cooled crystallization mixture may be filtered under vacuum, and theisolated solids may be washed with a suitable solvent, such as coldrecrystallization solvent, and dried under a nitrogen purge to affordthe desired crystalline form. The isolated solids may be analyzed by asuitable spectroscopic or analytical technique, such as solid statenuclear magnetic resonance, X-ray powder diffraction, or the like, toassure formation of the preferred crystalline form of the product. Theresulting crystalline form is typically produced in an amount of greaterthan about 70 weight percent isolated yield, preferably greater than 90weight percent isolated yield, based on the weight of the compoundoriginally employed in the crystallization procedure. The product may beco-milled or passed through a mesh screen to delump the product, ifnecessary.

Crystalline forms may be prepared, for example, directly from thereaction medium of the process for preparing Compound 2. This may beachieved, for example, by employing in the final process step a solventor a mixture of solvents from which Form N-1 may be crystallized.Alternatively, crystalline forms may be obtained by distillation orsolvent addition techniques. Suitable solvents for this purpose include,for example, non-polar solvents and polar solvents, including proticpolar solvents such as alcohols, and aprotic polar solvents such asketones, the details and selection of which are known to those skilledin the art.

The presence of more than one polymorph in a sample may be determined bytechniques such as powder X-ray diffraction (PXRD) or solid statenuclear magnetic resonance spectroscopy (SSNMR). For example, thepresence of extra peaks in an experimentally measured PXRD pattern whencompared with a simulated PXRD pattern may indicate more than onepolymorph in the sample. The simulated PXRD may be calculated fromsingle crystal X-ray data. see Smith, D. K., A FORTRAN Program forCalculating X-ray Powder Diffraction Patterns, Lawrence RadiationLaboratory, Livermore, Calif., UCRL-7196 (April 1963). In one aspect,Form N-1 has phase homogeneity indicated by less than 5 percent,preferably less than 2 percent, and more preferably less than 1 percentof the total peak area in the experimentally measured PXRD patternarising from the extra peaks that are absent from a simulated PXRDpattern.

Preferably, the crystallization technique provides a product consistingessentially of Form N-1. The crystallized material preferably comprisesat least 95 wt % of Form N-1, based on the weight of Compound 2 in thecomposition. The remaining material may comprise other form(s) of thecompound and/or reaction impurities and/or processing impurities arisingfrom its preparation. The presence of reaction impurities and/orprocessing impurities may be determined by analytical techniques knownin the art, such as, for example, chromatography, nuclear magneticresonance spectroscopy, mass spectrometry, or infrared spectroscopy.

Characterization

Form N-1 can be characterized using various techniques, which are wellknown to those of ordinary skill in the art. Examples ofcharacterization methods include, but are not limited to, single crystalX-ray diffraction, powder X-ray diffraction (PXRD), simulated powderX-ray patterns (Yin, S. et al., Am. Pharm. Rev., 6(2):80 (2003)),differential scanning calorimetry (DSC), solid-state ¹³C NMR (Earl, W.L. et al., J. Magn. Reson., 48:35-54 (1982)), Raman spectroscopy,infrared spectroscopy, moisture sorption isotherms, thermal gravimetricanalysis (TGA), and hot stage techniques.

The forms may be characterized and distinguished using single crystalX-ray diffraction, which is based on unit cell measurements of a singlecrystal of form N-1. A detailed description of unit cells is provided inStout et al., Ch. 3, X-ray Structure Determination: A Practical Guide,Macmillan Co., New York (1968), which is herein incorporated byreference. Alternatively, the unique arrangement of atoms in spatialrelation within the crystalline lattice may be characterized accordingto the observed fractional atomic coordinates. Another means ofcharacterizing the crystalline structure is by powder X-ray diffractionanalysis in which the diffraction profile is compared to a simulatedprofile representing pure powder material, both run at the sameanalytical temperature, and measurements for the subject formcharacterized as a series of 2θ values.

One of ordinary skill in the art will appreciate that an X-raydiffraction pattern may be obtained with a measurement of error that isdependent upon the measurement conditions employed. In particular, it isgenerally known that intensities in an X-ray diffraction pattern mayfluctuate depending upon measurement conditions employed. It should befurther understood that relative intensities may also vary dependingupon experimental conditions, and, accordingly, the exact order ofintensity should not be taken into account. Additionally, a measurementerror of diffraction angle for a conventional X-ray diffraction patternis typically about 5 percent or less, and such degree of measurementerror should be taken into account as pertaining to the aforementioneddiffraction angles. Consequently, it is to be understood that thecrystal forms of the present disclosure are not limited to the crystalforms that provide X-ray diffraction patterns completely identical tothe X-ray diffraction patterns depicted in the accompanying Figuresdisclosed herein. Any crystal form that provides an X-ray diffractionpattern, and DSC thermogram substantially identical to those disclosedin the accompanying Figures fall within the scope of the presentdisclosure. The ability to ascertain substantial identities of X-raydiffraction patterns is within the purview of one of ordinary skill inthe art.

Utility

Form N-1, alone or in combination with other compounds, can be used totreat depression. Form N-1, alone or in combination with othercompounds, can be used to treat other conditions, such as, for example,neurological conditions, psychiatric conditions and immunologicalconditions, e.g., anxiety disorders, pain, ADD, smoking cessation andobesity. Form N-1, alone or in combination with other compounds, i.e.,drugs, can be used to treat patients afflicted with various conditions(also referred to as “disorders”) by administering to said patients adose of a pharmaceutical composition provided herein. Examples ofdisorders that may be treatable by pharmaceutical compositionscomprising Form N-1 include, without limitation, ADD, attention deficitdisorder hyperactivity disorder (ADHD), cognition impairment, anxietydisorders, especially generalized anxiety disorder (GAD), panicdisorder, unipolar depression, also known as major depression, bipolardisorder, also known as manic depression or manic-depressive disorder,obsessive compulsive disorder (OCD), posttraumatic stress disorder(PTSD), acute stress disorder, social phobia, simple phobia,pre-menstrual dysphoric disorder (PMDD), social anxiety disorder (SAD),major depressive disorder (MDD), supranuclear palsy, eating disorders,especially obesity, anorexia nervosa, bulimia nervosa, and binge eatingdisorder, analgesia (including neuropathic pain, especially diabeticneuropathy), substance abuse disorders (including chemical dependencies)like nicotine addiction, cocaine addiction, alcohol and amphetamineaddiction, Lesch-Nyhan syndrome, neurodegenerative diseases likeParkinson's disease, late luteal phase syndrome or narcolepsy,psychiatric symptoms anger such as, rejection sensitivity, movementdisorders, like extrapyramidal syndrome, Tic disorders and restless legsyndrome (RLS), tardive dyskinesia, supranuclear palsy, sleep relatedeating disorder (SRED), night eating syndrome (NES), urinaryincontinence (including stress urinary incontinence (SUI) and mixedincontinence), migraine, fibromyalgia syndrome (FS), chronic fatiguesyndrome (CFS), sexual dysfunction especially premature ejaculation andmale impotence, thermoregulatory disorders (e.g., hot flashes that maybe associated with menopause), and lower back pain.

The present disclosure also provides pharmaceutical compositionscomprising a therapeutically effective amount of Form N-1 and at leastone pharmaceutically acceptable carrier.

The active ingredient, i.e., Form N-1 in such compositions typicallycomprises from 0.1 weight percent to 99.9 percent by weight of thecomposition, and often comprises from about 5 to 95 weight percent. Insome cases, the pH of the formulation may be adjusted withpharmaceutically acceptable modifiers (such as calcium carbonate andmagnesium oxide) to enhance the stability of the formulated compound orits delivery form. Formulations of the polymorph of the presentdisclosure may also contain additives for enhancement of absorption andbioavailability, or to improve the formulation process.

The pharmaceutical compositions of this disclosure may be administeredorally (as a solution or solid formulation), parenterally or via animplanted reservoir. The term parenteral as used herein includessubcutaneous, intracutaneous, intravenous, intramuscular,intra-articular, intrasynovial, intrasternal, intrathecal, andintralesional injection or infusion techniques.

The pharmaceutical compositions may be in the form of a sterileinjectable preparation, for example, as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according totechniques known in the art using suitable dispersing or wetting agentsand suspending agents. The details concerning the preparation of suchcompounds are known to those skilled in the art.

When orally administered, the pharmaceutical compositions of thisdisclosure may be administered in any orally acceptable dosage formincluding, but not limited to, capsules, tablets, and aqueoussuspensions and solutions. In the case of tablets for oral use, carrierswhich are commonly used include lactose and corn starch. Lubricatingagents, such as magnesium stearate, can also be added. For oraladministration in a capsule form, useful carriers/diluents includelactose, high and low molecular weight polyethylene glycol, and driedcorn starch. When aqueous suspensions are administered orally, theactive ingredient is combined with emulsifying and suspending agents. Ifdesired, certain sweetening and/or flavoring and/or coloring agents maybe added.

Other suitable carriers for the above noted compositions can be found instandard pharmaceutical texts, e.g., in Remington's PharmaceuticalSciences, 19th Ed., Mack Publishing Company, Easton, Pa. (1995). Furtherdetails concerning the design and preparation of suitable delivery formsof the pharmaceutical compositions of the disclosure are known to thoseskilled in the art.

As the skilled artisan will appreciate, the appropriate dosage of FormN-1 may be determined by a skilled medical practitioner. Specific dosageand treatment regimens for any particular patient will depend upon avariety of factors, including the activity of the specific compoundemployed, the age, body weight, general health status, gender, diet,time of administration, the duration of treatment, rate of excretion,drug combination, the severity and course of the condition, thepatient's disposition to the condition and the judgment of the treatingphysician.

The following non-limiting examples of reaction schemes that can be usedto prepare Form N-1. Synthesis schemes are illustrated below.

Preparation of Compound 2

Synthesis of compound 2 utilizes a general approach for C-7 and C-4substituted tetrahydroisoquinolines. A functional handle at C-7, allowsthe installation of the heterocycle using a palladium-catalyzed Suzukicoupling and the aryl substituent at C-4 is incorporated fromcommercially available starting materials. Although the route wasracemic, the first synthesis relied on separation of the racemic drugsubstance using in preparative chiral chromatography. The secondgeneration route relied on a classical resolution of an appropriateintermediate using di-p-toluoyl-D-tartaric acid (Synthesis Scheme 1).

The preferred synthesis begins with the reductive amination ofcommercially available m-anisealdehyde using methyl amine and sodiumbororhydride in an appropriate solvent such as a mixture of methanol andwater to afford compound 3 in quantitative yield (Synthesis Scheme 1).In this example the methoxy group will be the functional handle forinstallation of the C-7 pyridazine heterocycle. Alternativelym-bromobenzaldehyde has been demonstrated to be a suitable startingmaterial (Synthesis Scheme 2). A number of alternative reducing agentscan be envisioned to install the amine functionality Alkylation of thesecondary amine with α-bromo-2′-acetonapthone and an appropriate basecomposed of either an organic amine bases such as triethylamine,afforded ketone compound 4. Dichloromethane is the preferred solvent,although a number of solvents can be used including EtOAc, IPAC, MTBE,MeOH, EtOH, THF, ACN. Immediate reduction of the resulting ketone withsodium borohydride afforded compound 5 which was subsequently cyclizedunder acidic Friedel-Crafts conditions to afford racemictetrahydroisoquinoline compound 6 and its regioisomer in approximately2.5:1 ratio. A number of acids can be envisioned including protic acidssuch as methanesulfonic acid and sulfuric acid and Lewis acids such asTiCl₄ and AlCl₃. The regioisomers are separated via the oxlate salts andthen a selective crystallization in ethanol. The oxalate salt of thedesired isomer is then converted to the free base by treatment withaqueous sodium hydroxide and subsequently isolated by extraction withMTBE. A second iteration of this procedure improves the ration to >97:3.Classical resolution of the desired enantiomer, compound 7, is achievedusing di-p-toluoyl-D-tartaric acid. The desired tartrate salt is moreinsoluble in acetone, and isolated by filtration. After treatment withaqueous sodium hydroxide to afford the free base, the first pass affordsa 95:5 mixture of enantiomers. A second iteration provides >99% ee withchemical purity >99%. Alternatively the mixture of stereo- andregioisomers could be purified by chiral SFC chromatography.Demethylation of compound 7 with hydrobromic acid in acetic acid affordsthe phenol hydrobromide salt, compound 8, which is converted directly tothe triflate, compound 9, by using two equivalents of an organic aminebase, such as triethyl amine, DIPEA or pyridine, andtrifluoromethanesulfonic anhydride in dichloromethane. Alternatively thefreebase of compound 8 can be used successfully in the subsequent Suzukicoupling. A number of combinations of solvent, boronate, catalyst andligands can be envisioned for the subsequent Suzuki coupling. In thisexample, the crude triflate was converted to the crude boronate estercompound 10 using bis(pinacolato)diboron, KOAc and PdCl₂(dppf) in DMSO.Subsequent Suzuki coupling directly with 3-amino-6-chloropyridazineusing PdCl₂(dppf), Cs₂CO₃ in a DMF/water mixture afforded the desiredcompound 2 directly but required a tedious workup and laboriouspurification to afford pure product. Alternatively, usingN,N,-di-tert-butoxycarbonyl (Boc) protected 3-amino-6-chloropyridazine(compound 11) in the Suzuki coupling under similar conditions in amixture of DMSO and water afford compound 12. Compound 11 is readilyprepared from commercially available 3-amino-6-chloropyridazine,di-tert-butyl dicarbonate, and DMAP in DMF. Recrystallization of thebis-protected product from IPA/water significantly reduces the mono-bocamino pyridazine intermediate. Work-up of the crude reaction mixture ofcompound 12 with aqueous LiCl, aqueous NH₄OH, or treatment with asuitable metal scavengers such as Si-Thiol (SILICYCLE®), or activatedcarbon, or recrystallization from alcoholic solvent such as methanol areeffective means in reducing the metal contamination of the desiredproduct. The Boc protecting groups are removed under acidic conditionsusing HCl in an alcoholic solvent, such as methanol or isopropanol toafford the di-HCl salt, compound 13. The free base, compound 2, isproduced by crystallization from a mixture of aqueous sodium bicarbonateand methanol.

Alternatively on preparative scale: 3-amino-6-chloropyridazine can becoupled under Suzuki conditions with compound 18, to afford racemiccompound 1 (Synthesis Scheme 2). Compound 17, is readily accessibleusing similar procedures as discussed above and shown in SynthesisScheme 1, but starting with m-bromobenzaldehyde, where the C-7 bromineatom is the functional handle for installing the heterocycle. Separationof the desired (4S) enantiomer can be carried out on preparative scaleusing chiral HPLC on a CHIRALCEL® OD column. Subsequent treatment ofcompound 2 with L-tartaric acid in MeOH generated the L-tartrate salt,in quantitative yield.

Confirmation of Absolute Configuration of Compound 2 Via AsymmetricSynthesis of Compound 7

Proof of the absolute configuration of compound 2 was achieved byexecuting an asymmetric synthesis of compound 7 and X-ray analysis ofintermediate compound 22. (Synthesis Scheme 3). The asymmetric synthesisbegan with NBS-bromination of m-anisaldehyde to give compound 19 in 87%yield. The ketal compound 20 was prepared from compound 19 usingethylene glycol with camphorsulfonic acid as catalyst in quantitativeyield. Heck coupling of compound 20 with(R)-3-acryloyl-4-phenyloxazolidin-2-one using tri-o-tolyl phosphine andpalladium acetate gave compound 21 in 81% yield. Grignard addition ofthe complex formed by 2-naphthyl magnesium bromide and copper(I)bromide-dimethyl sulfide to a solution of compound 21 at −78° C., withwarming to room temperature, produced compound 22 in 85% yield, with ad.e. of approximately 98%. The absolute configuration of compound 22 wasconfirmed by single crystal X-ray crystallography (see structure below).The chiral auxiliary was cleaved by saponification to give carboxylicacid compound 23, which was converted with DPPA and MeOH to the methylcarbamate compound 24 in 59% yield. The ketal of compound 24 was cleavedat 0° C. with HCl. After isolation of the intermediate aldehyde, ringclosure under reductive amination conditions gave compound 25 in 60%yield. Reduction of the carbamate group using LiAlH₄ provided compound 7in 70% yield. This compound co-eluted by chiral LC with authenticmaterial prepared by the earlier methods.

EXAMPLES

The following examples are provided for illustrative purposes and arenot intended to limit the scope of the claimed invention.

Preparation of 1-(3-methoxyphenyl)-N-methylmethanamine (Compound 3)

To a solution of m-anisaldehyde (205 g, 1.5 mol) in methanol (800 mL) atroom temperature was added methylamine (130 mL of 40% in water, 1.5mol). The resulting solution was cooled to 0° C. and sodium borohydride(83 g, 2.3 mol) was added in batches. The reaction solution was stirredat 0° C. for 2 h, then warmed to room temperature, concentrated in vacuoand diluted with water. The organic phase was separated, diluted withethyl acetate, washed with 1:1 water/brine, dried over sodium sulfateand concentrated in vacuo to give the desired benzylamine (158.5 g, 70%)as a clear oil. The combined aqueous extracts were extracted withdichloromethane (3×). The combined methylene chloride extracts werewashed with 1:1 water/brine, dried over sodium sulfate and concentratedin vacuo to give additional benzylamine (66.7 g, 29%) as a clear oil: ¹HNMR (300 MHz, CDCl₃) 7.24 (t, J=8.4 Hz, 1H), 6.91-6.78 (m, 3H), 3.81 (s,3H), 3.73 (s, 2H), 2.46 (s, 3H).

Preparation of2-((3-methoxybenzyl)(methyl)amino)-1-(naphthalen-2-yl)ethanone (Compound4)

To a solution of α-bromo-2′-acetonaphthone (386.2 g, 1.57 mol) indichloromethane (2.5 L) at 0° C. was added compound 3 (249.1 g, 1.65mol) in 30 min, followed by the addition of triethylamine (220.7 mL,1.57 mmol) in 45 min. After stirring at 0° C. for 40 min, the reactionmixture was warmed to room temperature and stirred overnight. Thereaction solution was then washed with water (2×) and the aqueous layerwas re-extracted with dichloromethane (2×). The combined organic extractwas dried over sodium sulfate and concentrated in vacuo. The ketone,compound 4, was obtained as a reddish oil (513.3 g, quantitative), wasused in the next step without further purification: ¹H NMR (CDCl₃, 500MHz) 8.50 (s, 1H), 8.01 (d, J=8.5 Hz, 1H), 7.92 (d, J=8.0 Hz, 1H),7.88-7.85 (m, 2H), 7.60-7.54 (m, 2H), 7.26-7.23 (m, 1H), 6.97-6.94 (m,2H), 6.82 (dd, J=8.0, 2.5 Hz, 1H), 3.88 (s, 2H), 3.75 (s, 3H), 3.69 (s,2H), 2.42 (s, 3H); ESI MS m/z 320 [M+H]⁺.

Preparation of2-((3-methoxybenzyl)(methyl)amino)-1-(naphthalen-2-yl)ethanol (Compound5)

A solution of the ketone, compound 4, (512.2 g, 1.57 mol) in methanol(4.0 L) was split equally into two flasks. To each half of the ice-coldsolution was added sodium borohydride (33.0 g, 0.87 mol) in batches (−30min) After the addition, the reaction solution was stirred at 0° C.(internal temperature 10-15° C.) for 50 min before it was quenchedslowly with water (−500 mL). The reaction mixture was then concentratedin vacuo to remove most of the organic solvent. The residue obtainedfrom the two batches were combined, extracted with dichloromethane (2×),dried over sodium sulfate and concentrated in vacuo. The alcohol,compound 5, was obtained as a yellow oil (510.4 g, quantitative), andwas used in the next step without further purification: ¹H NMR (500 MHz,CDCl₃) δ 7.84-7.80 (m, 4H), 7.47-7.44 (m, 3H), 7.27-7.24 (m, 1H),6.92-6.83 (m, 3H), 4.92 (dd, J=10.0, 4.0 Hz, 1H), 4.14 (br s, 1H), 3.81(s, 3H), 3.74 (d, J=13.0 Hz, 1H), 3.53 (d, J=13.0 Hz, 1H), 2.68-2.60 (m,2H), 2.36 (s, 3H); ESI MS m/z 322 [M+H]⁺.

Preparation of racemic7-methoxy-2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinoline(Compound 6)

A solution of the alcohol, compound 5, (495 g, 1.54 mol) indichloromethane (6.0 L) was split equally into two flasks. To each halfof the ice-cold solution was added methanesulfonic acid (500 mL, 7.7mmol) via additional funnel (−1 h). The reaction solution was allowed toslowly warm to room temperature and stirred for 6-8 h before it wascooled to <10° C. and quenched slowly with an aqueous solution of sodiumhydroxide (330 g, 8.3 mol in 600 mL of water). The internal temperatureduring the addition was kept less than 30° C. The organic layer was thenseparated, washed with water, dried over sodium sulfate and concentratedin vacuo. The crude product from the two batches were combined andpurified by flash column chromatography (hexanes/ethyl acetate 88:12 toethyl acetate/methanol 99.9:0.1) to give compound 6 (280.4 g, 60% over 4steps) as a thick oil: ¹H NMR (500 MHz, CDCl₃) 7.81-7.74 (m, 3H), 7.68(s, 1H), 7.47-7.42 (m, 2H), 7.28-7.26 (m, 1H), 6.79 (d, J=8.5 Hz, 1H),6.80-6.62 (m, 2H), 4.40 (dd, J=8.5, 6.0 Hz, 1H), 3.80-3.78 (m, 1H), 3.77(s, 3H), 3.65 (d, J=15.0 Hz, 1H), 3.11-3.08 (m, 1H), 2.65 (dd, J=11.5,6.0 Hz, 1H), 2.45 (s, 3H); ESI MS m/z 304 [M+H]⁺.

Preparation of racemic7-methoxy-2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinoline(Compound 6)

Alternatively, the regioisomers can be separated as their respectiveoxalate salts. A 2.4:1 mixture of regioisomers [547 g, 1.8 mol] inabsolute EtOH (2 L) was stirred at ambient temperature as a solution ofoxalic acid (162 g, 1.8 mol, ACROS® lot #A0246832) in absolute EtOH (600mL) was added in one portion (exothermic). The solution becameheterogeneous, and after 2 h was filtered to give a light yellow solid[RBM-C-28(1)] that was ˜90:10 mixture of regioisomers by ¹H NMR. Thefilter cake was added to fresh absolute EtOH (6.5 L) and the resultingslurry was heated to 75° C. for 3 hours. The slurry was then cooled to25° C. and filtered. The filter cake was conditioned under N₂ overnightand then added to satd. NaHCO₃ (3 L). The product was extracted withEtOAc (3.5 L), and the organic layer was dried (MgSO₄) and concentratedto give 355 g of a white solid that was a 96:4 mixture of compound 6 andits regioisomer in 91% yield and 98.4% purity. ¹H NMR (400 MHz, CDCl₃) δ7.75 (m, 4H), 7.44 (m, 2H), 7.27 (dd, J=8.4, 1.7 Hz, 1H), 6.70 (m, 3H),4.39 (m, 1H), 3.77 (m, 4H), 3.63 (m, 1H), 3.08 (ddd, J=11.5, 4.3, 1.2Hz, 1H), 2.64 (dd, J=11.5, 8.9 Hz, 1H), 2.44 (s, 3H).

Preparation of(S)-7-methoxy-2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinoline(Compound 7)

To a solution of compound 6 (222.3 g, 0.73 mol) in ethanol (5.5 L) atroom temperature was quickly added a solution of di-p-toluoyl-D-tartaricacid (292.2 g, 0.73 mol) in ethanol (1.0 L) with no internal temperatureincrease noted. The reaction solution was stirred at room temperature.Precipitate started to form within 10 min. After stirring for 2 h, thereaction slurry was filtered and the cake was dried at 60° C. in vacuofor 12 h to give the (4S)-enriched tartrate salt (265.0 g, 52%, with aratio of enantiomers of approximately 84:16, CHIRALCEL® AD,heptane:IPA:diethyl amine 90:10:0.01). The filtrate was concentrated invacuo and dried at 60° C. to give the 4R-enriched tartrate salt (238.0g, 47%, ratio of enantiomers of approximately 3:97). The desired 4Senriched tartrate salt was treated with aqueous sodium hydroxide andseparated by preparative chiral chromatography to give pure compound 7.(Preparative chiral chromatography was carried out in two batches ofenriched racemate, 117 g and 93 g, respectively. The followingconditions were used for the 117 g batch: Thar 350 SFC; Column.CHIRALPAK® AD-H, 5×25 cm; Mobile phase: 30% IPA+0.05% DEA/CO₂; Pressure:100 bar; Temperature: 45° C.; Flow rate: 240 g/min; Solutionconcentration ˜250 mg/ml; Injection amount: 8 mL; Cycle time: 10.5min/inj; Detector: 254 nM; Throughput: 11-12 g/hr. The sample wasdissolved in warm 275 mL MeOH and 95 mL IPA with 0.5 mL DEA added).

Preparation of(S)-2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-ol(Compound 8, free base)

To a solution of compound 7 (42.5 g, 140 mmol) in 48% HBr (200 mL) wasadded AcOH (100 mL) to give a light yellow suspension. The reaction washeated to give a light yellow solution and stirred at 120° C. for 6 hr(<1.5% starting material left by HPLC), then concentrated.Dichloromethane (500 mL) was added to the residue and the suspension wasfiltered. The filtrate was neutralized with ice water, 50% NaOH (addedslowly) and Na₂CO₃ solution to pH of 10. The aqueous layer was extractedwith dichloromethane twice. The solid was fully dissolved indichloromethane and triethylamine (25 mL), and washed with Na₂CO₃solution to pH of 10. The organic layers were combined and washed withNaHCO₃ and brine, dried over Na₂SO₄ overnight, filtered andconcentrated. The crude product compound 8, as the free base, can beused in the next step without further purification.

Preparation of(S)-2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-ol(Compound 8, free base)

Alternative procedure: To a solution of compound 7 (30.0 g, 99.0 mmol)in acetic acid (150 mL) was added hydrobromic acid (48% solution inwater, 450 mL). The reaction solution was flushed with nitrogen andheated at 110° C. (internal temperature) for 3 h, at which time HPLC andMS showed no starting material left. The reaction solution was thencooled to room temperature and concentrated in vacuo. The residueobtained was dissolved in methylene chloride and water, then carefullyneutralized with saturated aqueous sodium bicarbonate and sodiumhydroxide to pH>8. The organic extract was separated, washed with waterand brine, dried over sodium sulfate and concentrated in vacuo to givecompound 8 as the freebase, (29.1 g, quantitative yield) as a lightyellow solid. The crude product was used in the next step withoutfurther purifications: ¹H NMR (500 MHz, CDCl₃) 7.81-7.74 (m, 3H), 7.67(s, 1H), 7.47-7.42 (m, 2H), 7.30-7.25 (m, 1H), 6.73 (d, J=8.5 Hz, 1H),6.57-6.52 (m, 2H), 4.38 (dd, J=8.5, 6.0 Hz, 1H), 3.73 (d, J=15.0 Hz,1H), 3.60 (d, J=15.0 Hz, 1H), 3.11-3.08 (m, 1H), 2.63 (dd, J=11.5, 9.5Hz, 1H), 2.44 (s, 3H); ESI MS m/z 290 [M+H]⁺.

Preparation of(S)-2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-ol(Compound 8, HBr salt)

Alternatively, the intermediate HBr salt of compound 8 can be isolatedand used directly in the next step.

Scaled-up procedure: A mixture of compound 7 (1.0 kg, 1.0 equiv,) inAcOH (2.5 L, 2.5 vol) was mechanically stirred and heated at 55-60° C.until the solution was homogeneous. 48 wt % HBr in water (5.0 L, 5.0vol) was then added in one portion, and the resulting solution washeated to 105° C. for 18 h (the solution became heterogeneous shortlyafter adding the HBr solution, but became homogeneous after stirringapproximately 2 h at 105° C.). The solution was then cooled to 95° C.over 15 minutes. DI water (900 mL) was added over 15 min, and then seedsof authentic Compound 26 (5.0 g) were added. An additional 3.1 L of DIwater was then added over 2 hours. The slurry was cooled to 25° C. over2.5 hours and filtered. The filter cake was conditioned under nitrogenovernight and then added to DI water (4.0 L). The slurry was stirred atambient temperature for 45 minutes, cooled to 5° C., filtered,conditioned under N₂ overnight, and then dried in vacuo to constantweight to give 1.0 kg of compound 8, HBr salt as a white solid in 85%yield and >99% purity. ¹H NMR (500 MHz, d-DMSO) δ ¹H NMR (500 MHz,d-DMSO) δ 10.31 (s, 0.85H), 10.11 (br s, 0.15H), 9.60 (s, 1H), 7.91 (m,4H), 7.54 (m, 2H), 7.30 (d, J=7.5 Hz, 1H), 6.67 (s, 1H), 6.61 (m, 2H),4.64 (m, 1H), 4.54 (br s, 2H), 3.84 (br s, 1H), 3.62 (t, J=11.2 Hz, 1H),2.98 (s, 3H).

Preparation of(S)-2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-yltrifluoromethanesulfonate (Compound 9)

To a solution of compound 8, free base (28.9 g, 99.0 mmol) indichloromethane (820 mL) was added pyridine (10.4 mL, 128.7 mmol). Thereaction suspension was stirred for 5 min at room temperature to give asolution. The reaction solution was cooled to 0° C. andtrifluoromethanesulfonic anhydride (18.5 mL, 108.9 mmol) was addedslowly (˜35 min). The reaction mixture was stirred at 0° C. for 45 minbefore it was quenched with aqueous sodium bicarbonate and extractedwith dichloromethane. The combined organic extract was dried over sodiumsulfate, filtered and concentrated in vacuo. Compound 9, obtained as ayellow oil, was used in the next step without further purification. ¹HNMR (500 MHz, CDCl₃) 7.83-7.70 (m, 3H), 7.67 (s, 1H), 7.49-7.44 (m, 2H),7.30-7.23 (m, 1H), 7.04 (d, J=2.05 Hz, 1H), 6.98-6.95 (m, 2H),), 4.42(dd, J=8.0, 6.0 Hz, 1H), 3.81 (dd, J=15.0 Hz, 1H), 3.68 (dd, J=15.0 Hz,1H), 3.11 (dd, J=11.0, 6.0 Hz, 1H), 2.69 (dd, J=11.0, 3.5 Hz, 1H), 2.46(s, 3H); ESI MS m/z 422 [M+H]⁺.

Preparation of(S)-2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-yltrifluoromethanesulfonate (Compound 9)

Alternative procedure: To a solution of compound 8, HBr salt (40.5 g,140 mmol) in DCM (1.2 L) was added diisopropylethylamine (48.9 mL, 280mmol) to give an orange solution. The reaction was cooled to −50° C.Triflic anhydride (Tf₂O) (35 mL, 207 mmol) was added in portions untilno starting material was left. The reaction was monitored by HPLC in 5min after each portion of Tf₂O addition. When the reaction was finished,aqueous NaHCO₃ solution (600 mL) was added. The organic layer was washedwith NH₄Cl solution (500 mL) and brine (500 mL). The combined organiclayer was dried over Na₂SO₄, filtered and concentrated. DMSO (500 mL)was added and the residual DCM was removed under reduced pressure togive crude product compound 9, which was used directly as is andcontinued in the example below. HPLC (YMC Pack Pro C18 4.6×50, 4 min):Peak at Tr=2.86 min is starting material. Peak at Tr=3.71 min isproduct.

Note: A test reaction indicated that the use of an excessive amount ofTf₂O may lead to demethylation and other undesired side reactions.

Preparation of(S)-2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-yltrifluoromethanesulfonate (Compound 9)

Scaled-up procedure: A mixture of compound 8 (1.0 kg, 1.0 equiv.) in DCM(12.4 L) was stirred at room temperature as Et₃N (0.90 L, 2.3 equiv) wasadded over 5 minutes (slightly exothermic). After stirring 20 minutes,the solution was homogeneous. The reactor was then cooled to −55 to −60°C. in a dry ice/IPA bath, and Tf₂O (0.56 L, 1.2 equiv.) was added over2.5 hours, maintaining an internal temperature of <−50° C. After theaddition was complete, the mixture was stirred for 20 minutes at −50° C.and HPLC analysis indicated the reaction was complete. The solution waswarmed to −10° C. and 10% aq. NaHCO₃ (6.2 L) was added in one portion.The resulting mixture was then warmed to 20° C. and stirred for 30minutes. The layers were then separated, and the organic phase waswashed with DI water (6.2 L) and concentrated under reduced pressure togive 1.2 kg of the crude product compound 9 as a dark red oil in >100%yield due to residual DCM, but with 73.5% purity.

Preparation of di-tert-butyl (6-chloropyridazin-3-yl)imidodicarbonate(Compound 11)

To a solution of 6-chloropyridazin-3-amine (75 g, 579 mmol) in DMF (600ml) was added di-tert-butyl dicarbonate (278 g, 1274 mmol) and DMAP (0.6g, 4.91 mmol) to give a suspension. The mixture was heated to 50° C. togive an orange solution and the reaction temperature reached 75° C. byitself with gas bubbles generated. The heating mantel was removed andthe reaction was cooled to 55° C. slowly. Then the reaction was stirredat 55° C. with heating for 2 h to give a dark brown solution. After thereaction was cooled to room temperature, the reaction solution waspoured to a 4 L beaker and water (3 L) was added to give a light brownsuspension. The yellow precipitate was collected by filtration andwashed with water (4 L). No product was dissolved in water. The solidwas dried in oven overnight to give product (152.2 g, 80% yield) as alight yellow solid.

Preparation of di-tert-butyl (6-chloropyridazin-3-yl)imidodicarbonate(Compound 11)

Scaled-up procedure: A mixture of 3-chloro-6-aminopyridazine (500 g, 1.0equiv) and DMAP (3.77 g, 0.008 equiv) in DMF (3.5 L) was heated andstirred at 55° C. for 20 minutes, until the mixture became homogeneous.A solution of Boc₂O (2.2 equiv) in DMF (500 mL) was prepared separatelyand then added in one portion to the batch, causing an endothermicevent. The reaction solution was monitored by HPLC while stirring at 55°C., and after 5 h, the reaction was deemed complete. The solution wascooled to room temperature and then slowly poured into DI water (4.4 L),causing precipitation. The mixture was stirred for 10 minutes and thenfiltered. The resulting brown solid was conditioned overnight undernitrogen. A mixture of the crude product in 2-propanol (3.5 L) washeated and stirred at 65° C. until the mixture became homogeneous. DIwater (3.5 L) was then added over 30 min (temperature was notcontrolled). The reaction mixture was cooled to room temperature andthen filtered to give 805 g compound 11 as a light brown solid in 63%yield, and 99.7% purity. ¹H NMR (300 MHz, d-DMSO) δ 8.00 (q, J=9.0 Hz,2H), 1.40 (s, 18H).

Preparation of(S)-di-tert-butyl-(6-(2-methyl-4-(2-naphthyl)-1,2,3,4-tetrahydro-7-isoquinolinyl)-3-pyridazinyl)imidodicarbonate(Compound 12)

(Continued from example above: To the solution of crude compound 9 (˜140mmol) (prepared above) in DMSO (800 mL) in a 5 L three-necked roundbottom flask was added bis(pinacolato)diboron (42.7 g, 168 mmol) andpotassium acetate (41.2 g, 420 mmol). The mixture was stirred at roomtemperature under nitrogen for 5 min followed by the addition ofdichloro[1,1′-ferrocenylbis(diphenyl-phosphine)]palladium(II)dichloromethane (Pd(dppf)Cl₂.CH₂Cl₂) (18.29 g, 22.40 mmol). The reactionwas stirred at 80° C. for 0.5 h. HPLC showed no starting materialremaining HPLC (YMC Pack Pro C18 4.6×50, 4 min): Peak at Tr=3.73 min isstarting material. Peaks at Tr=3.85 and 2.93 min are the productcompound 10 (LCMS=400.31 [M+H]) and boronic acid (LCMS=318.28 [M+H]),respectively.

To the solution of compound 10 prepared above was added compound 11 (175mmol), Pd(dppf)Cl₂.CH₂Cl₂) (18.29 g, 22.4 mmol), DMSO (400 mL), Cs₂CO₃(137 g, 420 mmol) and water (250 mL) at 80° C. The reaction was stirredat 80° C. for 2 h. The reaction was cooled to room temperature anddiluted with water (6 L). The black precipitate was collected byfiltration, dissolved in DCM (1 L), and washed with 10% LiCl aqueoussolution (500 mL). The aqueous layer was back extracted with DCM (400mL). The combined organic layers were filtered through a CELITE® pad andconcentrated. Crude product compound 12 was obtained as a dark greysolid (57.4 g, 71% yield, 98% purity) by crystallization in MeOH (300ml). HPLC (YMC Pack Pro C18 4.6×50, 4 min): Peak at Tr=3.80 is theproduct compound 12. LCMS=567.33 [M+H].

Purification of(S)—N,N-di-tert-butyl-(6-(2-methyl-4-(2-naphthyl)-1,2,3,4-tetrahydro-7-isoquinolinyl)-3-pyridazinyl)imidodicarbonate(Compound 12)

1. Several batches was combined together for purification. To a solutionof crude compound 12 (120 g, 98% purity, dark grey solid) indichloromethane (1 L) was added activated carbon (50 g). The suspensionwas refluxed for 1 h and the charcoal was filtered off through a CELITE®pad. The filtrate was concentrated and the residue was recrystallized inMeOH (1 L, reflux 30 min). The solution was cooled to room temperatureand the grey crystals were collected by filtration. Therecrystallization was repeated two times.

2. The solid was de-colored with charcoal in dichloromethane andrecrystallized in MeOH again to give a beige solid.

3. The solid was dissolved in dichloromethane (1 L) and washed withammonium hydroxide solution (100 ml concentrated NH₄OH+900 ml water) toprimarily remove residual iron. The middle black suspension layerbetween the aqueous layer and dichloromethane layer was removed with theuse of a separation funnel. The light yellow dichloromethane layer wasdried over Na₂SO₄ and concentrated. The residue was recrystallized inMeOH (500 ml) again to give off-white color solid.

4. The solid was dissolved in dichloromethane (500 ml) and the solutionwas concentrated. The residue was recrystallized in MeOH (500 ml) andthe product was collected by filtration as a white solid. Firstfraction, 95 g, >99% purity, white solid; second fraction, 10 g, 98%purity from last MeOH mother liquid; third fraction, 1 g, 90% purityfrom former mother liquid. ¹H NMR (500 MHz, CDCl₃) 7.96 (s, 1H),7.66-7.89 (m, 6H), 7.40-7.53 (m, 3H), 7.28 (d, J=7.15 Hz, 1H), 7.04 (d,J=8.25 Hz, 1H), 4.46-4.56 (m, 1H), 3.84 (dd, J=80.56, 15.12 Hz, 2H),3.15 (dd, J=11.55, 6.05 Hz, 1H), 2.71 (dd, J=11.55, 8.80 Hz, 1H), 2.49(s, 3H), 1.46 (s, 18H). ¹³C NMR (126 MHz, CDCl₃) ppm 157.78, 154.91,150.59, 141.51, 139.60, 136.19, 133.48, 132.47, 130.24, 128.18, 127.84,127.67, 127.63, 127.13, 126.08, 125.77, 125.63, 125.17, 125.07, 125.01,83.92, 61.43, 58.44, 46.00, 45.90, 27.88.

Preparation of(S)—N,N-di-tert-butyl-(6-(2-methyl-4-(2-naphthyl)-1,2,3,4-tetrahydro-7-isoquinolinyl)-3-pyridazinyl)imidodicarbonate(Compound 12)

Scaled-up procedure: A solution of compound 9 (1.18 kg, 1.0 equiv) inDMSO (16.5 L, 14 vol) was stirred at ambient temperature in a 50-Ljacketed reactor as bis-pinacolato diboron (0.85 kg, 1.2 equiv) and KOAc(0.82 kg, 3.0 equiv) were added. The heterogeneous mixture was spargedwith a strong stream of nitrogen for 1.5 hours. PdCl₂(dppf) (60 g, 0.025equiv) was then added and the mixture was heated and stirred at 85° C.under nitrogen. After 5 hours, HPLC analysis indicated the reaction hadstalled at ˜67% conversion, so an additional 10 g of PdCl₂(dppf) wasadded. After 2 h, the reaction had again stalled at 75% conversion. Thesolution was sparged with N₂ for 30 min, and sparging was continuedthroughout the rest of the reaction. An additional 10 g of PdCl₂(dppf)was added, and the reaction was complete within 2.5 h to afford areaction mixture containing compound 10. A solution of Cs₂CO₃ (2.74 kg,3.0 equiv) in DI water (4.2 L) that had been sparged with a strongstream of N₂ for 1.5 hours was then added in one portion. A slurry ofcompound 11 (1.15 kg, 1.25 equiv) in DMSO (6.5 L) that had been spargedwith a strong stream of N₂ for 1.5 hours was then added in one portion(delayed exotherm). A final charge of PdCl₂(dppf) (60 g) was added, andthe reaction mixture was stirred at 85° C. for 12 h and then cooled to25° C. DI water (5.0 L) was added, and after stirring for 1 hour, themixture was filtered to give a brown solid, which was conditioned underN₂ overnight.

The filter cake was dissolved in DCM (18 L) and the organic layer waswashed with a 10% aqueous LiCl solution (18 L) and a 10% aqueous NH₄OHsolution (18 L). The DCM layer was concentrated under reduced pressureto leave a brown solid. MeOH (6 L) was added and the slurry was stirredand heated at 55° C. for 1 hour. It was then cooled to 25° C. and EtOAc(12 L) was added. The resulting slurry was stirred for 1 hour, filtered,and the filter cake was conditioned under N₂ overnight to give 750 g ofa grey solid. ICP analysis indicated the palladium content was ˜2300ppm. The solid was then dissolved in DCM (3.75 L) and Si-thiol (1.50 kg,2 wt equiv, SILICYCLE® lot #10347) was added. The mixture was vigorouslystirred at 30-35° C. for 4.5 hours. The mixture was cooled to roomtemperature and then filtered. The solid Si-thiol was then rinsed withDCM (7.5 L) and the combined dark brown filtrates were transferred to aRotovap bulb through two 1.2 micron filters and concentrated underreduced pressure to give an off-white solid. ICP analysis indicated thepalladium content was 160 ppm. The batch was re-dissolved in DCM (3.75L) and Si-thiol (1.50 kg, 2 wt equiv) was added. After stirring for 4.5hours at 30-35° C., the mixture was cooled to room temperature andfiltered. The solid Si-thiol was rinsed with DCM (7.5 L), and thecombined yellow filtrates were transferred to a Rotovap bulb through a1.2 micron filter and concentrated under reduced pressure to give anoff-white solid. ICP analysis indicated the palladium content was 3 ppm.EtOAc (4.2 L) was then added to the solid and the resulting slurry wasstirred at ambient temperature for 1 hour and then filtered. The filtercake was rinsed with EtOAc (500 mL). The filter cake was conditionedunder N₂ for 2 days to give 715 g of a compound 12 as a white solid in47% yield and >99% purity. ¹H NMR (500 MHz, d-DMSO) δ 8.31 (d, J=9.1 Hz,1H), 8.02 (d, J=1.3 Hz, 1H), 7.87 (m, 5H), 7.79 (s, 1H), 7.48 (m, 2H),7.39 (dd, J=8.5, 1.5 Hz, 1H), 7.01 (d, J=8.2 Hz, 1H), 4.49 (t, J=6.3 Hz,1H), 3.78 (m, 2H), 3.02 (dd, J=11.5, 6.6 Hz, 1H), 2.73 (dd, J=11.4, 7.1Hz, 1H), 2.39 (s, 3H), 1.41 (s, 18H).

Preparation of(S)-2-methyl-4-(naphthalen-2-yl)-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroisoquinoline(Compound 10)

Alternative procedure: To a mixture of the triflate compound 9 (99.0mmol), bis(pinacolato)diboron (30.2 g, 118.8 mmol) and potassium acetate(29.1 g, 297.0 mmol) was added DMSO (725 mL). The resulting solution waspurged with argon for 10 min, and thendichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II)dichloromethane adduct (6.47 g, 7.92 mmol) was added. The reactionsolution was degassed again with argon for 5 min, heated at 82° C. (oilbath) for 1 h and then cooled to room temperature and poured into water(1.0 L). The mixture was extracted with ethyl acetate (800 mL) and theorganic extract was separated and washed with water (2×) and brine,dried over sodium sulfate and concentrated in vacuo. The crude boronateester compound 10 (48.0 g), obtained as a brown foam, was used in thenext step without further purification: ESI MS m/z 400 [M+H]⁺.

Preparation of(S)-6-(2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-yl)pyridazin-3-amine(Compound 2)

Alternative procedure directly from 3-amino-6-chloropyridazine: To amixture of the boronate ester compound 10 (23.0 g, 47.4 mmol),3-amino-6-chloropyridazine (9.2 g, 71.1 mmol) and cesium carbonate (46.3g, 142.2 mmol) were added DMF (464 mL) and water (116 mL). The reactionsolution was flushed with argon for 10 min, and thendichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II)dichloromethane adduct (3.9 g, 4.74 mmol) was added. The mixture wasflushed with argon for 5 min and heated at 80° C. for 1 h. The reactionsolution was cooled to room temperature, diluted with ethyl acetate,washed with 1:1 brine and water (2×), dried over sodium sulfate andconcentrated in vacuo. The crude product was purified by flash columnchromatography (99:0.9:0.1 to 94:5.4:0.6 ethylacetate/methanol/concentrated ammonium hydroxide). This partiallypurified product (14.8 g as a brown solid) was stirred with ethylacetate (120 mL) under argon for 12 h and filtered to give compound 2(12.5 g, 72%) as a brown solid: ¹H NMR (500 MHz, CDCl₃) 7.82-7.76 (m,4H), 7.71 (s, 1H), 7.61-7.57 (m, 2H), 7.47-7.43 (m, 2H), 7.30 (dd,J=8.0, 1.5 Hz, 1H), 6.99 (d, J=8.0 Hz, 1H), 6.80 (d, J=9.5 Hz, 1H), 4.71(br s, 2H), 4.50 (t, J=7.0 Hz, 1H), 3.89 (d, J=15.0 Hz, 1H), 3.74 (d,J=14.5 Hz, 1H), 3.14 (dd, J=11.5, 6.0 Hz, 1H), 2.70 (dd, J=11.5, 9.0 Hz,1H), 2.48 (s, 3H); ESI MS m/z 367 [M+H]⁺.

Preparation of(S)-6-(2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-yl)pyridazin-3-aminedihydrochloride (Compound 13)

Compound 12 (193 g, 341 mmol) was dissolved in 2.5 M hydrochloride inethanol (1088 ml, 2720 mmol). The solution was heated at 75° C. for 1 h.The resulted suspension was cooled to room temperature and diluted withEtOAc (2 L). The white solid was collected by filtration, washed withEtOAc (1 L). The white solid was dried at 40° C. overnight under reducedpressure to give compound 13 (143 g, 96% yield, 99.8% purity): ¹H NMR(500 MHz, DMSO-d₆) ppm: 8.77 (s-broad, 2H); 8.37 (d, J=9.71 Hz, 1H);7.95 (overlap, 1H); 7.94 (m, 2H); 7.94 (overlap, 1H); 7.91 (s, 1H); 7.76(d, J=8.01 Hz, 1H); 7.66 (d, J=9.71 Hz, 1H); 7.56 (m, 1H); 7.55 (m, 1H);7.33 (broad, 1H); 6.94 (broad, 1H); 4.92 (m, 1H); 4.66 (s-broad, 2H);3.85 (m, 1H); 3.73 (m, 1H); 2.97 (s, 3H). HRMS [M+H]⁺=367.19202.

Preparation of(S)-6-(2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-yl)pyridazin-3-amine(Compound 2)

A solution of compound 13 (30.0 g, 0.68 mmol,) in 1:1 (v/v) MeOH/H₂O(900 mL) was filtered through a glass-sintered funnel. The filtrate wasthen stirred at room temperature as a saturated solution of aqueousNaHCO₃ (45 mL) was slowly added over 30 minutes, causing the solution toturn cloudy. Previously prepared seed crystals (0.10 g,) were then addedto the batch in one portion. Additional saturated aqueous NaHCO₃ (555mL) was added to the reaction mixture over 1.5 h. The slurry wasfiltered to afford a wet cake. The wet filter cake was then suspended in8:2 (v/v) H₂O/MeOH (250 mL) and the resulting slurry was stirred for 3hours and then filtered. The filter cake was washed with 8:2 (v/v)H₂O/MeOH (100 mL) and the obtained white solid was dried in vacuo at 50°C. for 30 h to give compound 2 (23.2 g, 93% yield): ¹H NMR (d₆-DMSO, 300MHz) 7.80-7.90 (m, 3H), 7.74-7.78 (m, 3H), 7.65 (dd, J=8.6; 2.1 Hz, 1H),7.43-7.53 (m, 2H), 7.38 (dd, J=8.6; 2.1 Hz, 1H), 6.90 (d, J=8.0 Hz, 1H),6.83 (d, J=9.1 Hz, 1H), 6.45 (s, 2H), 4.43 (t, J=5.9 Hz, 1H), 3.73 (s,2H), 3.00 (dd, J=10.7, 5.4 Hz, 1H), 2.70 (dd, J=11.3; 7.5 Hz, 1H), 2.37(s, 3H); ¹³C NMR (75 MHz, DMSO) ppm 159.69, 149.65, 142.55, 136.95,135.62, 134.79, 132.93, 131.83, 129.47, 127.67, 127.46, 127.40, 127.28,127.16, 125.98, 125.47, 125.15, 123.32, 123.06, 114.12, 60.63, 57.82,45.55, 44.69, 40.34.

Preparation of(S)-6-(2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-yl)pyridazin-3-aminedihydrochloride (Compound 13)

Scaled-up procedure: A mixture of compound 12 (750 g, 1.0 equiv) in IPA(3.75 L) was stirred at room temperature as 5-6 N HCl in IPA (3.75 L,)was added over 5 minutes (slightly exothermic). The mixture was thenheated to 70° C. and held for 18 h, during which time the mixture becamea homogeneous yellow solution, and then a white slurry. HPLC analysisindicated the reaction was complete. The mixture was cooled to 25° C.and EtOAc (8 L) was then added in one portion. After stirring for 1hour, the mixture was filtered to give 555 g of a compound 13 as a whitesolid in 95% yield, and with >99% purity. ¹H NMR (500 MHz, d-DMSO) δ12.05 (br s, 1H), 8.83 (br s, 2H), 8.38 (d, J=9.7 Hz, 1H), 7.94 (m, 5H),7.76 (d, J=8.1 Hz, 1H), 7.69 (d, J=9.7 Hz, 1H), 7.55 (m, 2H), 7.33 (s,1H), 6.93 (d, J=5.3 Hz, 1H), 4.92 (m, 1H), 4.68 (br s, 2H), 3.85 (m,1H), 3.76 (m, 1H), 3.55 (br s, 1H), 2.97 (s, 3H).

Preparation of(S)-6-(2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-yl)pyridazin-3-aminedihydrochloride (Compound 13)

Alternative procedure for preparation of compound 13, bis-HCl salt, fromcompound 2, Form N-1 free base: To a solution of compound 2 (35.9 g,98.0 mmol) in ethanol (750 mL) at room temperature was addedconcentrated HCl (28.7 mL) via additional funnel. The solution washeated at 80° C. and additional EtOH (500 mL) and water (55 mL) wereadded. After all the additions, the reaction mixture was stirred at 80°C. for 30 min upon which all solids dissolved. The solution was thenfiltered and the filtrate was allowed to cool to room temperature andstand overnight. The precipitate formed was collected by filtration anddried at 40° C. in vacuo for 5 h to give the desired bis-HCl saltcompound 13 (28.7 g) as an off-white solid. ¹H NMR (500 MHz, DMSO-d₆)ppm: 8.77 (s-broad, 2H); 8.37 (d, J=9.71 Hz, 1H); 7.95 (overlap, 1H);7.94 (m, 2H); 7.94 (overlap, 1H); 7.91 (s, 1H); 7.76 (d, J=8.01 Hz, 1H);7.66 (d, J=9.71 Hz, 1H); 7.56 (m, 1H); 7.55 (m, 1H); 7.33 (broad, 1H);6.94 (broad, 1H); 4.92 (m, 1H); 4.66 (s-broad, 2H); 3.85 (m, 1H); 3.73(m, 1H); 2.97 (s, 3H). HRMS [M+H]E=367.19202.

Preparation of(S)-6-(2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-yl)pyridazin-3-amine(Compound 2)

A solution of compound 13 (550 g, 1.0 equiv) in 1:1 MeOH/DI water (16.5L) was passed through a 1.2 micron in-line filter. The solution wasstirred at ambient temperature as a 10% aq. NaHCO₃ solution was slowlyadded. The batch became cloudy after 350 mL of the NaHCO₃ solution hadbeen added. Seed crystals (10.5 g) were then added, and the mixture wasstirred for an additional 1 hour. More of the NaHCO₃ solution (4 L) wasadded over 40 minutes. After the addition was complete, the mixture wasstirred for 2 hours and then filtered. After conditioning on the filterunder N₂ for 1 hour, the wet filter cake was re-suspended in 8:2 DIwater/MeOH (5.5 L), stirred for 2 hours, and then filtered. The filtercake was conditioned under N₂ for 48 hours and then dried in vacuo at35° C. for 48 hours to give 450 g of the compound 2 as a white solid in96% yield and >99% purity.

Examples for Synthesis Scheme 3: Asymmetric synthesis of Compound 7Preparation of 2-bromo-5-methoxybenzaldehyde (Compound 19))

To a solution of m-anisaldehyde (55.4 g, 0.41 mol) in DMF (400 mL) wasadded a solution of N-bromosuccinimide (124.0 g, 0.69 mol) dropwise atroom temperature. After the addition, the reaction solution was stirredat room temperature for 12 h, then poured into a mixture of ice andwater and stirred for 10 min. The precipitate was collected byfiltration and dissolved in ethyl acetate. The resulting solution waswashed with water (2×) and brine, dried over sodium sulfate andconcentrated in vacuo to give compound 19 (76.4 g, 87%) as an off-whitesolid: ¹H NMR (CDCl₃, 500 MHz) 10.32 (s, 1H), 7.53 (d, J=8.5 Hz, 1H),7.42 (d, J=3.5 Hz, 1H), 7.04 (dd, J=9.0, 3.0 Hz, 1H), 3.85 (s, 3H).

Preparation of 2-(2-bromo-5-methoxyphenyl)-1,3-dioxolane (Compound 20)

To a solution of compound 19 (50.0 g, 0.23 mol) in toluene (650 mL) wereadded ethylene glycol (14.2 mL, 0.26 mol) and camphorsulfonic acid (10.7g, 46 mmol). The reaction solution was heated under reflux with aDean-Stark trap for 6 h and then cooled to room temperature and dilutedwith ethyl acetate (300 mL). The resulting solution was washed withaqueous saturated sodium bicarbonate and brine, dried over sodiumsulfate and concentrated in vacuo to give compound 20 (61.5 g,quantitative) as a yellow oil: ¹H NMR (CDCl₃, 500 MHz) 7.44 (d, J=8.5Hz, 1H), 7.15 (d, J=3.5 Hz, 1H), 6.79 (dd, J=8.5, 3.0 Hz, 1H), 6.04 (s,1H), 4.18-4.06 (m, 4H), 3.81 (s, 3H).

Preparation of(R,E)-3-(3-(2-(1,3-dioxolan-2-yl)-4-methoxyphenyl)acryloyl)-4-phenyloxazolidin-2-one(Compound 21)

A mixture of compound 20 (2.6 g, 10.0 mmol),(R)-3-acryloyl-4-phenyloxazolidin-2-one (2.2 g, 10.0 mmol), tri-o-tolylphosphine (0.30 g, 1.0 mmol) and palladium acetate (0.11 g, 0.5 mmol) intriethylamine (35 mL) under argon was stirred under reflux for 90 min.The resulting reaction mixture was cooled to room temperature andconcentrated in vacuo. The residue obtained was purified by flash columnchromatography (hexanes/ethyl acetate 95:5 to 60:40) to give compound 21(3.2 g, 81%) as a light yellow oil: ¹H NMR (500 MHz, CDCl₃) 8.19 (d,J=15.5 Hz, 1H), 7.81 (d, J=15.5 Hz, 1H), 7.40 (d, J=8.5 Hz, 1H),7.41-7.30 (m, 5H), 7.17 (d, J=3.0 Hz, 1H), 6.91 (dd, J=8.5, 3.0 Hz, 1H),6.01 (s, 1H), 5.54 (dd, J=9.0, 3.5 Hz, 1H), 4.73 (t, J=9.0 Hz, 1H), 4.30(dd, J=9.0, 4.0 Hz, 1H), 4.32-4.28 (m, 2H), 4.13-4.10 (m, 2H), 3.85 (s,3H).

Preparation of(R)-3-((S)-3-(2-(1,3-dioxolan-2-yl)-4-methoxyphenyl)-3-(naphthalen-2-yl)propanoyl)-4-phenyloxazolidin-2-one(Compound 22)

To a solution of copper(I) bromide-dimethyl sulfide complex (1.43 g, 6.9mmol) in anhydrous THF (33 mL) and dimethyl sulfide (16.5 mL) at −78° C.was added 2-naphthylene magnesium bromide (27.3 mL, 0.5 M in THF) slowlyvia syringe. After the addition, the reaction solution was stirred at−40° C. for 30 min, and then re-cooled to −78° C. A solution of Compound21 (1.8 g, 4.6 mmol) in THF (24 mL) was then added via syringe. Thereaction mixture was slowly warmed to room temperature before it wasquenched with aqueous ammonium chloride at 0° C. and extracted withethyl acetate (2×). The combined organic extract was washed with 1:1water/brine, dried over sodium sulfate and concentrated in vacuo. Theresidue obtained was purified by flash column chromatography(hexanes/ethyl acetate 95:5 to 55:45) to give compound 22 (2.15 g, 85%)as an off-white foam: ¹H NMR (CDCl₃, 500 MHz) 7.72-7.70 (m, 2H),7.69-7.63 (m, 2H), 7.46-7.40 (m, 2H), 7.37 (d, J=8.5 Hz, 1H), 7.24 (d,J=8.5 Hz, 1H), 7.17-7.11 (m, 2H), 7.04 (t, J=7.5 Hz, 2H), 6.99 (d, J=7.5Hz, 2H), 6.81 (s, J=8.5 Hz, 1H), 6.09 (s, 1H), 5.34-5.30 (m, 2H), 4.59(t, J=8.5 Hz, 1H), 4.16-4.14 (m, 3H), 4.06-4.01 (m, 3H), 3.79 (s, 3H),3.60 (dd, J=16.5, 8.5 Hz, 1H).

Preparation of(S)-3-(2-(1,3-dioxolan-2-yl)-4-methoxyphenyl)-3-(naphthalen-2-yl)propanoicacid (Compound 23)

To a solution of compound 22 (0.55 g, 1.0 mmol) in a mixture of THF (12mL) and water (4 mL) at 0° C. was added hydrogen peroxide (0.41 mL, 30%in water), followed by lithium hydroxide (48 mg, 2.0 mmol) in water (2.5mL). The reaction solution was stirred at 0° C. for 1 h and roomtemperature for 30 min. A solution of sodium sulfite (0.78 g) in water(5 mL) was then added. After stirring at 0° C. for 10 min, the mixturewas concentrated in vacuo to remove the organic solvent. The remainingaqueous solution was diluted with aqueous sodium hydroxide and extractedwith dichloromethane. The basic aqueous layer was separated, neutralizedwith aqueous ammonium chloride to pH 6-7 and then extracted withdichloromethane (3×). The combined organic extract was dried over sodiumsulfate and concentrated in vacuo to give compound 23 (0.41 g, crude),which was used in the next step without further purification: ¹H NMR(500 MHz, CDCl₃) δ 7.56-7.70 (m, 4H), 7.48-7.33 (m, 3H), 7.13 (d, J=2.5Hz, 1H), 7.08 (d, J=8.5 Hz, 1H), 6.80 (dd, J=9.0, 3.0 Hz, 1H), 6.08 (s,1H), 5.18 (dd, J=9.0, 7.0 Hz, 1H), 4.21-4.00 (m, 4H), 3.77 (s, 3H), 3.20(dd, J=16.0, 9.0 Hz, 1H), 3.12 (dd, J=16.0, 7.0 Hz, 1H).

Preparation of (S)-methyl2-(2-(1,3-dioxolan-2-yl)-4-methoxyphenyl)-2-(naphthalen-2-yl)ethylcarbamate(Compound 24)

To a solution of compound 23 (0.38 g, 1.0 mmol) and triethylamine (0.15mL, 1.0 mmol) in toluene (10 mL) at 85-90° C. was addeddiphenylphosphorylazide (0.21 mL, 1.0 mmol) via syringe. The reactionmixture was stirred for 30 min, then it was cooled to 50° C. andmethanol (0.30 mL, 7.5 mmol) was added to it. The resultant solution wasstirred at 50° C. for 14 h, then cooled to room temperature, dilutedwith ethyl acetate and washed with aqueous ammonium chloride. Theorganic extract was dried over sodium sulfate and concentrated in vacuo.Purification by flash column chromatography (hexanes/ethyl acetate 95:5to 50:50) gave compound 24 (0.24 g, 59%) as a white foam: ¹H NMR (500MHz, CDCl₃) 7.81-7.72 (m, 4H), 7.48-7.42 (m, 2H), 7.33 (d, J=8.5 Hz,1H), 7.17-7.14 (m, 2H), 6.84 (dd, J=8.5, 2.5 Hz, 1H), 6.05 (s, 1H), 5.22(br s, 1H), 4.81 (t, J=8.0 Hz, 1H), 4.22-4.07 (m, 4H), 3.98-3.87 (m,2H), 3.79 (s, 3H), 3.71-3.52 (m, 3H).

Preparation of (S)-methyl7-methoxy-4-(naphthalen-2-yl)-3,4-dihydroisoquinoline-2(1H)-carboxylate(Compound 25)

To a solution of compound 24 (0.10 g, 0.25 mmol) in 1,4-dioxane (10 mL)at 0° C. was added concentrated HCl (0.8 mL). The reaction solution wasstirred at room temperature for 20 min, and then quenched with aqueoussodium bicarbonate and extracted with dichloromethane (2×). The combinedorganic extract was dried over sodium sulfate and concentrated in vacuo.The residue obtained was dissolved in a mixture of ethanol (30 mL) andtrifluoroacetic acid (2 mL). Palladium on carbon (105 mg) was added andthe reaction mixture was shaken under hydrogen (25 psi) for 15 min, thenfiltered through a pad of CELITE® and concentrated in vacuo. The residuewas dissolved in dichloromethane and the resulting solution was washedwith aqueous sodium bicarbonate, dried over sodium sulfate andconcentrated in vacuo. Purification by preparative thin layerchromatography (hexanes/ethyl acetate 75:25) provided compound 25 (52mg, 60%) as a colorless oil: ¹H NMR (500 MHz, CDCl₃) 7.82-7.74 (m, 3H),7.62-7.51 (m, 1H), 7.47-7.44 (m, 2H), 7.24 (d, J=9.0 Hz, 1H), 6.85 (d,J=8.5 Hz, 1H), 6.80-6.68 (m, 2H), 4.83-4.60 (m, 2H), 4.40-3.90 (m, 2H),3.81 (s, 3H), 3.76-3.42 (m, 4H); ESI MS m/z 348 [M+H]⁺.

Preparation of(S)-7-methoxy-2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinoline(Compound 7)

To a solution of compound 25 (51 mg, 0.15 mmol) in THF (8 mL) at 0° C.was added lithium aluminum hydride (0.6 mL, 1M in THF) dropwise. Afterthe addition, the reaction solution was heated under reflux for 1 h,cooled to 0° C. and quenched by sequential addition of water (1.8 mL),aqueous sodium hydroxide (0.6 mL) and water (0.6 mL). The resultantmixture was stirred at 0° C. for 10 min before it was filtered. Thefiltrate was extracted with dichloromethane (2×). The combined organicextract was dried over sodium sulfate and concentrated in vacuo.Purification by preparative thin layer chromatography(dichloromethane/methanol/concentrated ammonia 95:4.5:0.5) providedcompound 7 (32.0 mg, 70%, AUC HPLC 96.8%, CHIRALPAK® AD 100%) as a lightyellow oil: [α]²³ _(D)+50.7° (c 0.18, methanol); ¹H NMR (500 MHz, CDCl₃)7.79-7.68 (m, 3H), 7.68 (s, 1H), 7.47-7.43 (m, 2H), 7.28-7.26 (m, 1H),6.80 (d, J=8.0 Hz, 1H), 6.65-6.62 (m, 2H), 4.39 (dd, J=8.5, 6.0 Hz, 1H),3.78 (s, 3H), 3.77 (d, J=15.0 Hz, 1H), 3.65 (d, J=14.5 Hz, 1H), 3.09(dd, J=11.5, 5.5 Hz, 1H), 2.64 (dd, J=11.5, 9.0 Hz, 1H), 2.44 (s, 3H);ESI MS m/z 304 [M+H]⁺. This compound co-elutes with authentic compound 7on CHIRALPAK® AD (90:10:0.1 heptane:IPA:diethyl amine, 1 mL/min, RT=14.3min. The RT of the (−)-enantiomer of compound 7=6.2 min).

Preparation of 1-(3-bromophenyl)-N-methylmethanamine (Compound 14)

To a solution of 3-bromobenzaldehyde (138 g, 0.74 mol) in methanol (0.9L) was added a 40% aqueous solution of methylamine (64 mL, 0.82 mol)followed by stirring at 0° C. for 1 h. Sodium borohydride (42.3 g, 1.1mol) was added in portions at 0° C. and the reaction mixture was stirredovernight while warming to room temperature. The solution wasconcentrated, then diluted with water (300 mL). The resulting solutionwas extracted with methylene chloride (3×300 mL) and chloroform (2×300mL). The combined organic extracts were washed with brine (2×200 mL)then dried over sodium sulfate, filtered and concentrated under reducedpressure to afford crude compound 14 (148 g) as a clear oil, which wasused in the next step without further purification: ¹H NMR (CDCl₃, 300MHz). 7.48 (s, 1H), 7.39-7.34 (m, 1H), 7.27-7.16 (m, 2H), 3.71 (s, 2H),2.43 (s, 3H), 1.38 (s, 1H).

Preparation of2-((3-bromobenzyl)(methyl)amino)-1-(naphthalen-2-yl)ethanone (Compound15)

To a solution of compound 14 (23.1 g, 115.5 mmol) in methylene chloride(500 mL) was added α-bromo-2′-acetonapthone (27.9 g, 110.0 mmol) and theresulting mixture was stirred at 0° C. for 1 h prior to the addition oftriethylamine (15.3 mL, 47.4 mmol). The reaction mixture was stirred at0° C. for 2 h. The resulting mixture was diluted with water (200 mL) andthe aqueous phase was extracted with additional methylene chloride(2×200 mL). The combined extracts were dried over sodium sulfate,filtered and concentrated to afford compound 15 (44.5 g) as a lightyellow oil, which was used in the next step without furtherpurification: ¹H NMR (CDCl₃, 500 MHz) 8.49 (s, 1H), 8.01 (dd, J=8.8, 1.8Hz, 1H), 7.95 (d, J=8.0 Hz, 1H), 7.90-7.84 (m, 2H), 7.62-7.52 (m, 3H),7.40 (d, J=8.0 Hz, 1H), 7.31 (d, J=8.0 Hz, 1H), 7.19 (t, J=7.7 Hz, 1H),3.91 (s, 2H), 3.68 (s, 2H), 2.40 (s, 3H).

Preparation of2-((3-bromobenzyl)(methyl)amino)-1-(naphthalen-2-yl)ethanol (Compound16)

To a solution of compound 15 (−110 mmol) in methanol (600 mL), sodiumborohydride (5.4 g, 142.8 mmol) was added in portions at 0° C. Thereaction mixture was first stirred overnight while warming up to roomtemperature. The reaction mixture was concentrated under reducedpressure. The residue was diluted with water (200 mL) and the solutionwas extracted with methylene chloride (3×300 mL). The combined organicextracts were dried over sodium sulfate, filtered and concentrated todryness under reduced pressure to afford the crude desired productcompound 16 (42.6 g) as a yellow oil, which was used without furtherpurification in the next step: ¹H NMR (300 MHz, CDCl₃) δ 7.86-7.79 (m,4H), 7.52-7.38 (m, 5H), 7.28-7.15 (m, 2H), 4.93 (dd, J=9.5, 4.4 Hz, 1H),4.05 (s, 1H), 3.71 (d, J=13.3 Hz, 1H), 3.52 (d, J=13.3 Hz, 1H),2.72-2.61 (m, 2H), 2.34 (s, 3H).

Preparation of7-bromo-2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinoline(Compound 17)

To a solution of compound 16 (−110 mmol) in methylene chloride (1.0 L)was added concentrated sulfuric acid (30.0 mL, 0.56 mol) and the mixturewas stirred at 0° C. for 3 h. The reaction was quenched by adding 6 NNaOH until the pH was ˜9, and the aqueous phase was extracted withadditional methylene chloride (3×). The combined organic extracts weredried over sodium sulfate, filtered and concentrated. The residue waspurified by flash chromatography (9:1 to 8:1 hexanes/ethyl acetate) toafford compound 17 (15.79 g, 41% over 3 steps) as a white solid: ¹H NMR(500 MHz, CDCl₃) 7.82-7.70 (m, 3H), 7.66 (s, 1H), 7.49-7.41 (m, 2H),7.28-7.20 (m, 2H), 7.16 (dd, J=8.3, 2.3 Hz, 1H), 6.75 (d, J=8.5 Hz, 1H),4.37 (dd, J=7.5, 6.5 Hz, 1H), 3.75 (d, J=15.3 Hz, 1H), 3.62 (d, J=15.3Hz, 1H), 3.08 (ddd, J=11.5, 6.0, 1.0 Hz, 1H), 2.64 (dd, J=11.5, 8.5 Hz,1H), 2.43 (s, 3H). The undesired 5-bromo isomer was also obtained (11.91g, 30% over 3 steps).

Preparation of2-methyl-4-(naphthalen-2-yl)-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroisoquinoline(Compound 18)

To a mixture of compound 17 (19.3 mmol), bis(pinacolato)diboron (5.9 g,23.2 mmol), and potassium acetate (5.7 g, 57.9 mmol) was added DMSO (140mL). The resultant solution was purged with argon for 10 min, and thendichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II)dichloromethane adduct (1.3 g, 1.5 mmol) was added. The reactionsolution was degassed again with argon for 5 min, then heated at 80° C.(oil bath) for 1 h. The resultant solution was cooled to roomtemperature, diluted with ethyl acetate and washed with water and brine.The organic layer was separated, dried over sodium sulfate andconcentrated under reduced pressure. The crude boronate ester2-methyl-4-(naphthalen-2-yl)-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroisoquinolinecompound 18 (10.0 g), obtained as a brown foam, was used in the nextstep without further purification: ESI MS m/z 400 [M+H]⁺.

Preparation ofrac-6-(2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-yl)pyridazin-3-amine(rac-1)

To a mixture of compound 18 (6.0 g, 15.0 mmol),3-amino-6-chloropyridazine (3.0 g, 22.5 mmol) and cesium carbonate (14.7g, 45.0 mmol) were added DMF (140 mL) and water (35 mL). The reactionsolution was flushed with argon for 10 min, and thendichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II)dichloromethane adduct (1.2 g, 1.5 mmol) was added. The resultantmixture was flushed with argon for 5 min and heated at 80° C. for 1 h.The reaction solution was then cooled to room temperature, diluted withethyl acetate, washed with 1:1 brine and water (2×), dried over sodiumsulfate and concentrated in vacuo. The crude product was purified byflash column chromatography (92:2:5.4:0.6 to 47:47:5.4:0.6dichloromethane/ethyl acetate/methanol/concentrated ammonium hydroxide)to give rac-1 (3.2 g, 58%) as a light tan solid: ¹H NMR (500 MHz, CDCl₃)7.82-7.76 (m, 4H), 7.71 (s, 1H), 7.61-7.57 (m, 2H), 7.47-7.43 (m, 2H),7.30 (dd, J=8.0, 1.5 Hz, 1H), 6.99 (d, J=8.0 Hz, 1H), 6.80 (d, J=9.5 Hz,1H), 4.71 (br s, 2H), 4.50 (t, J=7.0 Hz, 1H), 3.89 (d, J=15.0 Hz, 1H),3.74 (d, J=14.5 Hz, 1H), 3.14 (dd, J=11.5, 6.0 Hz, 1H), 2.70 (dd,J=11.5, 9.0 Hz, 1H), 2.48 (s, 3H); ESI MS m/z 367 [M+H]⁺.

Preparation of(S)-6-(2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-yl)pyridazin-3-amine(Compound 2)

Rac-1 (3.3 g) was resolved by preparative chiral HPLC (CHIRALCEL® ODcolumn, using 80:20:0.1 heptane/ethanol/diethylamine as the eluent) togive the (S)-enantiomer compound 2 [[α]²⁵ _(D)+122.0° (c 0.15,methanol)] (1.6 g) as a light brown foam and the (R)-enantiomer [[α]²⁵_(D)−124.3° (c 0.23, methanol)] (1.6 g) as an off-white solid.

Preparation of(S)-6-(2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-yl)pyridazin-3-amine,L-tartrate (Compound 2, L-tartrate)

To a mixture of compound 2, free base (1.6 g, 4.3 mmol) and L-tartaricacid (0.65 g, 4.3 mmol) were added methanol (280 mL) and water (100 mL).The reaction slurry was sonicated and heated at approximately 55° C. togive a clear solution, which was then concentrated in vacuo to about 100mL. The resulting solution was lyophilized to give compound 2-L-tartrate(2.18 g, 98.6%, AUC HPLC >99%) as an off-white solid: mp 153-158° C.; ¹HNMR (500 MHz, CDCl₃) 7.87-7.80 (m, 6H), 7.69 (dd, J=8.0, 1.5 Hz, 1H),7.51-7.48 (m, 2H), 7.30 (dd, J=8.5, 1.5 Hz, 1H), 7.04-7.01 (m, 2H), 4.71(dd, J=11.0, 5.5 Hz, 1H), 4.43 (s, 2H), 4.39 (d, J=16.0 Hz, 1H), 4.29(d, J=15.5 Hz, 1H), 3.64 (dd, J=11.5, 5.5 Hz, 1H), 3.37-3.30 (m, 1H),2.85 (s, 3H); ESI MS m/z 367 [M+H]⁺; Anal. Calcd. ForC₂₄H₂₂N₄.C₄H₆O₆.H₂O: C, 62.91; H, 5.66; N, 10.48. Found: C, 62.81; H,5.73; N, 10.30.

Preparation of(S)-6-(2-methyl-4-(naphthalen-2-yl)-1,2,3,4-tetrahydroisoquinolin-7-yl)pyridazin-3-amine,L-tartrate (Compound 2, L-tartrate salt)

Alternative procedure: To a slurry of compound 2, free base (1.5 g, 4.1mmol) in methanol (80 mL) was added a solution of L-tartaric acid (0.63g, 4.2 mmol) in methanol (15 mL) and water (10 mL). The reaction slurrybecame a clear solution after heating under reflux. The resultingsolution was then cooled to 0° C. while stirring and precipitationoccurred. The precipitate obtained was collected by filtration. Thissolid was re-crystallized twice from methanol (150 mL) and water (20mL). The resulting solid was dissolved in methanol (25 mL) and water(100 mL), and the solution was lyophilized to give the L-tartrate saltof compound 2, (1.4 g, 65%, AUC HPLC >99%) as a white solid: [α]²³_(D)+79.1° (c 0.15, methanol); ¹H NMR (500 MHz, CDCl₃) 7.87-7.80 (m,6H), 7.69 (dd, J=8.0, 1.5 Hz, 1H), 7.51-7.48 (m, 2H), 7.30 (dd, J=8.5,1.5 Hz, 1H), 7.04-7.01 (m, 2H), 4.71 (dd, J=11.0, 5.5 Hz, 1H), 4.43 (s,2H), 4.39 (d, J=16.0 Hz, 1H), 4.29 (d, J=15.5 Hz, 1H), 3.64 (dd, J=11.5,5.5 Hz, 1H), 3.37-3.30 (m, 1H), 2.85 (s, 3H); ESI MS m/z 367 [M+H]⁺.

Form N-1 was analyzed using one or more of the testing methods describedbelow.

Single Crystal X-Ray Measurements

A Nonius Kappa CCD diffractometer equipped with graphite-monochromatedMo Kα radiation (λ=0.7107 Å) was used to collect diffraction data at theroom temperature (Nonius 2001a. Data Collection Software for NoniusKappa-CCD devices. Nonius BV, Delft, The Netherlands. Nonius 2001b;DENZO Processing Software for Nonius Kappa-CCD devices, Nonius BV,Delft, The Netherlands). The final unit cell parameters were determinedusing the entire data set.

All structures were solved by direct methods and refined by thefull-matrix least-squares techniques, using the SHELXTL software package(Sheldrick, G. M. (1997), SHELXTL. Structure Determination Programs.Version 5.10, Bruker AXS, Madison, Wis., USA.). The function minimizedin the refinements was Σ_(W)(|F_(O)|−|F_(C)|)². R is defined asΣ∥F_(O)|−|F_(C)∥/Σ|F_(O)| whileR_(W)=[Σ_(W)(|F_(O)|−|F_(C)|)₂/Σ_(W)|F_(O)|²]^(1/2), where w is anappropriate weighting function based on errors in the observedintensities. Difference Fourier maps were examined at all stages ofrefinement. All non-hydrogen atoms were refined with anisotropic thermaldisplacement parameters. The hydrogen atoms associated with hydrogenbonding were located in the final difference Fourier maps while thepositions of the other hydrogen atoms were calculated from an idealizedgeometry with standard bond lengths and angles. They were assignedisotropic temperature factors and included in structure factorcalculations with fixed parameters.

The crystal data of the N-1 form is shown in Table 2. The fractionalatomic coordinates are listed in Table 3. It should be understood by oneof ordinary skill in the art that slight variations in the coordinatesare possible and are considered to be within the scope the presentdisclosure.

TABLE 2 Crystal Data of Form N-1 Temperature room temperature Wavelength0.71073 Å Crystal system, space group Monoclinic, P2₁ Unit celldimensions a = 8.4299(4) Å alpha = 90° b = 6.0698(3) Å beta =100.169(2)° c = 19.0689(12) Å gamma = 90° Volume 960.39(9) Å³ Z,Calculated density 2, 1.267 g/cm³

TABLE 3 Atomic Coordinates x y z U(eq) N1 0.3248(4) −0.7759(7)−0.0061(2) 0.0608(10) N2 0.3353(4) −0.4151(6) 0.0322(2) 0.0516(9) N30.2832(4) −0.2519(6) 0.07049(19) 0.0504(8) N4 −0.2362(4) 0.4479(6)0.20586(19) 0.0525(9) C1 0.2608(5) −0.6106(7) 0.0275(2) 0.0476(9) C20.1165(5) −0.6421(7) 0.0547(2) 0.0476(9) C3 0.0665(5) −0.4763(7)0.0935(2) 0.0487(9) C4 0.1596(4) −0.2835(6) 0.1041(2) 0.0439(8) C50.1240(5) −0.1073(6) 0.1527(2) 0.0453(9) C6 −0.0334(4) −0.0444(6)0.1549(2) 0.0447(9) C7 −0.0674(4) 0.1306(6) 0.1973(2) 0.0446(9) C8−0.2368(5) 0.2120(8) 0.1916(3) 0.0556(10) C9 −0.3995(6) 0.5360(11)0.1966(4) 0.0778(15) C10 −0.1522(5) 0.4871(7) 0.2779(2) 0.0541(10) C110.0289(4) 0.4359(6) 0.2862(2) 0.0463(9) C12 0.0594(4) 0.2382(7)0.2416(2) 0.0441(8) C13 0.2163(5) 0.1695(7) 0.2403(2) 0.0541(11) C140.2491(5) 0.0035(8) 0.1957(2) 0.0528(11) C15 0.1013(4) 0.4074(6)0.3638(2) 0.0439(8) C16 0.1983(5) 0.5661(6) 0.4007(2) 0.0445(9) C170.2648(4) 0.5393(6) 0.4738(2) 0.0450(9) C18 0.3664(5) 0.6998(7)0.5124(2) 0.0533(10) C19 0.4321(6) 0.6659(9) 0.5822(3) 0.0681(13) C200.4004(6) 0.4740(9) 0.6174(2) 0.0661(13) C21 0.3015(5) 0.3166(8)0.5816(3) 0.0611(11) C22 0.2296(4) 0.3441(7) 0.5095(2) 0.0449(9) C230.1271(5) 0.1861(7) 0.4707(2) 0.0529(10) C24 0.0654(5) 0.2157(7)0.4011(2) 0.0527(10) H1A 0.4107 −0.7535 −0.0236 0.073 H1B 0.2796 −0.9034−0.0099 0.073 H16 0.2208 0.6937 0.3773 0.053 H11 0.0810 0.5636 0.26840.056 H6 −0.1181 −0.1205 0.1275 0.054 H14 0.3554 −0.0346 0.1945 0.063H24 −0.0020 0.1086 0.3769 0.063 H3 −0.0272 −0.4903 0.1126 0.058 H130.3012 0.2375 0.2703 0.065 H19 0.4991 0.7727 0.6068 0.082 H2 0.0579−0.7721 0.0463 0.057 H21 0.2812 0.1886 0.6054 0.073 H20 0.4462 0.45310.6650 0.079 H10A −0.1992 0.3957 0.3106 0.065 H10B −0.1662 0.6399 0.29040.065 H23 0.1014 0.0591 0.4936 0.063 H8A −0.2964 0.1828 0.1442 0.067 H8B−0.2897 0.1348 0.2257 0.067 H18 0.3887 0.8294 0.4900 0.064 H9A −0.45380.4762 0.2325 0.117 H9B −0.4572 0.4962 0.1503 0.117 H9C −0.3952 0.69360.2008 0.117

Powder X-Ray Diffraction

X-ray powder diffraction (PXRD) data were obtained using a Bruker C2GADDS. The radiation was Cu Kα (40 KV, 40 mA). The sample-detectordistance was 15 cm. Powder samples were placed in sealed glasscapillaries of 1 mm or less in diameter; the capillary was rotatedduring data collection. Data were collected for 3≦2θ≦35° with a sampleexposure time of at least 1000 seconds. The resulting two-dimensionaldiffraction arcs were integrated to create a traditional 1-dimensionalPXRD.

The results of the PXRD pattern and a simulated pattern calculated fromthe single crystal data are shown in FIG. 1.

Table 4 lists the characteristic PXRD peaks that describe Form N-1 ofCompound 2.

TABLE 4 Characteristic diffraction peak positions (degrees 2θ ± 0.1) atroom temperature, based on a high quality pattern collected with adiffractometer (cuKα) with a spinning capillary with 2θ calibrated witha NIST other suitable standard N-1 4.6 9.4 10.6 14.1 15.4 18.2 19.5

Differential Scanning Calorimetry

Differential scanning calorimetry (DSC) experiments were performed in aTA INSTRUMENTS° model Q1000 or 2920. The sample (about 2-6 mg) wasweighed in a pinpricked hermetically sealed aluminum pan and recordedaccurately recorded to a hundredth of a milligram, and transferred tothe DSC. The instrument was purged with nitrogen gas at 50 mL/min. Datawere collected between room temperature and 300° C. at 10° C./minheating rate. The plot was made with the endothermic peaks pointingdown.

The results are shown in FIG. 2.

Solid-State Nuclear Magnetic Resonance (SSNMR)

All solid-state C-13 NMR measurements were made with a Bruker DSX-400,400 MHz NMR spectrometer. High resolution spectra were obtained usinghigh-power proton decoupling and the TPPM pulse sequence and rampamplitude cross-polarization (RAMP-CP) with magic-angle spinning (MAS)at approximately 12 kHz (Bennett, A. E. et al., J. Chem. Phys., 103:6951(1995); Metz, G. et al., J. Magn. Reson. A, 110:219-227 (1994)).Approximately 70 mg of sample, packed into a canister-design zirconiarotor was used for each experiment. Chemical shifts (δ) were referencedto external adamantane with the high frequency resonance being set to38.56 ppm (Earl, W. L. et al., J. Magn. Reson., 48:35-54 (1982)).

The results are shown in FIG. 3.

In a preferred aspect, Form N-1 has the properties set for the in Table5, below.

TABLE 5 Physical and Chemical Properties Chemical name6-[(4S)-1,2,3,4-tetrahydro-2-methyl-4-(2-naphthalenyl)-7-isoquinolinyl]-3- pyridazinamine Chemical structure

Molecular formula C₂₄H₂₂N₄ Molecular weight 366.46 Appearance White tooff-white powder Melting point/range 237°-243° C. Solution pH ~7.0 atabout 1 μg/mL concentration in water pH-Solubility At solution pH 6.5and above, the aqueous profile solubility of Compound 2 is ~1 μg orless. Aqueous solubility increases at lower pH values (15.6 mg/mL at pH2.0 and 1.76 mg/mL at pH 4.4) Solubility profile practically insoluble:n-heptane (USP definition) very slightly soluble: acetonitrile, ethylacetate, n-butanol slightly soluble: isopropanol, acetone, ethanol,methanol, propylene glycol, dichloromethane sparingly soluble: PEG 400soluble: N,N-dimethylacetamide, dimethylsulfoxide freely soluble:tetrahydrofuran pK_(a) 4.9 and 7.8 Distribution Log D_(o/b) = 3.10 at pH6.5 and 3.82 at pH 7.4 coefficient Stability Compound 2 is stable up to25° C. with protection from light.

The examples and characterizations provided above are not intended tolimit the scope of the invention. For example, Form N-1 may be useful inthe form of the free base or acid or in the form of a pharmaceuticallyacceptable salt thereof. Similarly, it may be desirable to utilize aprodrug of form N-1.

What is claimed is:
 1. Form N-1 of


2. Form N-1 of

characterized by the following unit cell parameters: Cell dimensions:a=8.4299(4) Å b=6.0698(3) Å c=19.0689(12) Å alpha=90° beta=100.169(2)°gamma=90° Space group Monoclinic, P2₁ Volume 960.39(9) Å³ Z, Calculateddensity 2, 1.267 g/cm³ wherein measurement of said crystalline form isat a temperature between about 20° C. to about 25° C.
 3. Form N-1 of

characterized by fractional atomic coordinates within the unit cell aslisted in Table
 3. 4. Form N-1 of

with characteristic peaks in the powder X-ray diffraction pattern atvalues of two theta of 4.6±0.1, 9.4±0.1, 10.6±0.1, 14.1±0.1, 15.4±0.1,18.2±0.1, 19.5±0.1 at a temperature between about 20° C. and about 25°C.
 5. Form N-1 of

characterized by a melt with decomposition endotherm with onsettypically in the range of 235-245° C.
 6. Substantially pure Form N-1 of


7. The form of claim 6 wherein said Form N-1 has a purity of at least 98weight percent.
 8. The form of claim 6 wherein said Form N-1 has apurity of at least 99 weight percent.
 9. A pharmaceutical compositioncomprising Form N-1 of

and a pharmaceutically acceptable carrier.
 10. A pharmaceuticalcomposition comprising Form N-1 of

in combination with one or more additional drugs.
 11. A method oftreating depression in a mammal comprising administering to the mammal atherapeutically-effective amount of Form N-1 of


12. The method of claim 11 wherein the mammal is a human.
 13. A methodof treating anxiety disorders in a mammal comprising administering tothe mammal a therapeutically-effective amount of Form N-1 of


14. A method of treating pain in a mammal comprising administering tothe mammal a therapeutically-effective amount of Form N-1 of


15. A method of treating obesity in a mammal comprising administering tothe mammal a therapeutically-effective amount of Form N-1 of